IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Coronavirus nsp10/nsp16 Methyltransferase Can Be Targeted by nsp10-Derived Peptide In Vitro and In Vivo To Reduce Replication and Pathogenesis. Coronavirus nsp10/nsp16 Methyltransferase Can Be Targeted by nsp10-Derived Peptide In Vitro and In Vivo To Reduce Replication and Pathogenesis. Journal of virology 2015 imex curation Wang Y., Sun Y., Wu A., Xu S., Pan R., Zeng C., Jin X., Ge X., Shi Z., Ahola T., Chen Y., Guo D. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus Uniprot identifiers entered here are based on reviewed entries with the highest sequence identities to the stated Genbank accession numbers. chenyu@whu.edu.cn,dguo@whu.edu.cn 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro methyltransferase as methyltransferase assay methyltransferase as predetermined predetermined participant predetermined Wang Y., Sun Y., Wu A., Xu S., Pan R., Zeng C., Jin X., Ge X., Shi Z., Ahola T., Chen Y., Guo D. Virus - Publications including interactions involving viral proteins Reaction mixtures were spotted onto TLC plates. Radiolabelling was determined using a PhosphorImager. (see ref. 28 pmid:22022266). Uniprot identifiers entered here are based on reviewed entries with the highest sequence identities to the stated Genbank accession numbers. chenyu@whu.edu.cn,dguo@whu.edu.cn Accepted 2020-JUL-27 AT 14:47 BST AT 14:47 BST by PPORRAS Journal of virology 2015 Only protein-protein interactions imex curation imex curation Coronavirus nsp10/nsp16 Methyltransferase Can Be Targeted by nsp10-Derived Peptide In Vitro and In Vivo To Reduce Replication and Pathogenesis. Coronavirus nsp10/nsp16 Methyltransferase Can Be Targeted by nsp10-Derived Peptide In Vitro and In Vivo To Reduce Replication and Pathogenesis. Journal of virology 2015 imex curation Wang Y., Sun Y., Wu A., Xu S., Pan R., Zeng C., Jin X., Ge X., Shi Z., Ahola T., Chen Y., Guo D. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus Uniprot identifiers entered here are based on reviewed entries with the highest sequence identities to the stated Genbank accession numbers. chenyu@whu.edu.cn,dguo@whu.edu.cn 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro methyltransferase as methyltransferase assay methyltransferase as tag visualisation tag visualisation Wang Y., Sun Y., Wu A., Xu S., Pan R., Zeng C., Jin X., Ge X., Shi Z., Ahola T., Chen Y., Guo D. Virus - Publications including interactions involving viral proteins nsp10 and nsp16 of the various coronaviruses were incubated with the substrate m7GpppA-RNA in the presence of [3H]AdoMet. Samples were then analyzed by liquid scintillation assays after purification of the 3H-labeled substrates with DEAE-Sephadex chromatography. Uniprot identifiers entered here are based on reviewed entries with the highest sequence identities to the stated Genbank accession numbers. chenyu@whu.edu.cn,dguo@whu.edu.cn Accepted 2020-JUL-27 AT 14:47 BST AT 14:47 BST by PPORRAS Journal of virology 2015 Only protein-protein interactions imex curation imex curation p0c6y5-pro_0000037400 Putative 2'-O-methyl transferase rep ORF1ab polyprotein 1a-1b protein protein TGEV Porcine transmissible gastroenteritis coronavirus (strain Purdue) SAEWNPGYSMPTLYKIQRMCLERCNLYNYGAQVKLPDGITTNVVKYTQLCQYLNTTTLCVPHKMRVLHLGAAGASGVAPGSTVLRRWLPDDAILVDNDLRDYVSDADFSVTGDCTSLYIEDKFDLLVSDLYDGSTKSIDGENTSKDGFFTYINGFIKEKLSLGGSVAIKITEFSWNKDLYELIQRFEYWTVFCTSVNTSSSEGFLIGINYLGPYCDKAIVDGNIMHANYIFWRNSTIMALSHNSVLDTPKFKCRCNNALIVNLKEKELNEMVIGLLRKGKLLIRNNGKLLNFGNHFVNTP 6385 6684 0D2EEC28B1F0070F p0c6y5-pro_0000037395 Non-structural protein 10 rep ORF1ab polyprotein 1a-1b protein protein TGEV Porcine transmissible gastroenteritis coronavirus (strain Purdue) AGKPTEHPSNSSLLTLCAFSPDPAKAYVDAVKRGMQPVNNCVKMLSNGAGNGMAVTNGVEANTQQDSYGGASVCIYCRCHVEHPAIDGLCRYKGKFVQIPTGTQDPIRFCIENEVCVVCGCWLNNGCMCDRTSMQ 3864 3998 607DDF78BA90E58D p0c6y0-pro_0000037363 Non-structural protein 10 rep ORF1ab polyprotein 1a-1b protein protein MHV-JHM Murine coronavirus (strain JHM) Murine hepatitis virus AGTATEYASNSAILSLCAFSVDPKKTYLDYIQQGGVPVTNCVKMLCDHAGTGMAITIKPEATTNQDSYGGASVCIYCRSRVEHPDVDGLCKLRGKFVQVPLGIKDPVSYVLTHDVCQVCGFWRDGSCSCVGTGSQFQ 4324 4460 915B3D43CB036B21 q98vg9-pro_5000140217 Non-structural protein 10 rep ORF1ab polyprotein 1a-1b protein protein FCoV Feline coronavirus (strain FIPV WSU-79/1146) AGKPTEHPSNSGLLTLCAFAPDPAKAYVDAVKRGMQPVTNCVKMLSNGAGNGMAITNGVESNTQQDSYGGASVCIYCRCHVEHPAIDGLCRFKGKFVQVPTGTQDPIRFCIENEVCVVCGCWLTNGCMCDRTSIQ 3889 4023 62A581CCA720FA27 pppac20-rna-substrate pppAC20 RNA substrate rna ribonucleic acid RNA rna synthetic construct synthetic construct ACCCCCCCCCCCCCCCCCCCC mce_vaccw Cap-specific mRNA (nucleoside-2'-O-)-methyltransferase PAPS Poly(A) polymerase regulatory subunit Poly(A) polymerase small subunit VP39 J3R VACWR095 protein protein vaccw vaccinia virus (stra Vaccinia virus (strain WR) VACV MDVVSLDKPFMYFEEIDNELDYEPESANEVAKKLPYQGQLKLLLGELFFLSKLQRHGILDGATVVYIGSAPGTHIRYLRDHFYNLGVIIKWMLIDGRHHDPILNGLRDVTLVTRFVDEEYLRSIKKQLHPSKIILISDVRSKRGGNEPSTADLLSNYALQNVMISILNPVASSLKWRCPFPDQWIKDFYIPHGNKMLQPFAPSYSAEMRLLSIYTGENMRLTRVTKSDAVNYEKKMYYLNKIVRNKVVVNFDYPNQEYDYFHMYFMLRTVYCNKTFPTTKAKVLFLQQSIFRFLNIPTTSTEKVSHEPIQRKISSKNSMSKNRNSKRSVRSNK 55A299DA4AAE1AD4 55A299DA4AAE1AD4 k9n7c7-pro_0000422448 Non-structural protein 10 rep ORF1ab polyprotein 1a-1b protein protein cvemc Middle East respiratory syndrome-related coronavirus (isolate United Kingdom/H123990006/2012) (Betacoronavirus England 1) Betacoronavirus England 1 Human coronavirus EMC AGSNTEFASNSSVLSLVNFTVDPQKAYLDFVNAGGAPLTNCVKMLTPKTGTGIAISVKPESTADQETYGGASVCLYCRAHIEHPDVSGVCKYKGKFVQIPAQCVRDPVGFCLSNTPCNVCQYWIGYGCNCDSLRQAALPQ 4238 4377 7EE46B6A26B7A272 p0c6x7-pro_0000037322 2'-O-methyltransferase nsp16 ORF1ab polyprotein 1a-1b rep protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus ASQAWQPGVAMPNLYKMQRMLLEKCDLQNYGENAVIPKGIMMNVAKYTQLCQYLNTLTLAVPYNMRVIHFGAGSDKGVAPGTAVLRQWLPTGTLLVDSDLNDFVSDADSTLIGDCATVHTANKWDLIISDMYDPRTKHVTKENDSKEGFFTYLCGFIKQKLALGGSIAVKITEHSWNADLYKLMGHFSWWTAFVTNVNASSSEAFLIGANYLGKPKEQIDGYTMHANYIFWRNTNPIQLSSYSLFDMSKFPLKLRGTAVMSLKENQINDMIYSLLEKGRLIIRENNRVVVSSDILVNN 6776 7073 14B6660399D3A801 p0c6y1-pro_0000037412 Non-structural protein 10 rep ORF1ab polyprotein 1a-1b protein protein ibvb ibvb IBV SKGHETEEVDAVGILSLCSFAVDPADTYCKYVAAGNQPLGNCVKMLTVHNGSGFAITSKPSPTPDQDSYGGASVCLYCRAHIAHPGSVGNLDGRCQFKGSFVQIPTTEKDPVGFCLRNKVCTVCQCWIGYGCQCDSLRQPKSSVQ 3784 3928 297665FA9F44AC49 k9n7c7-pro_0000422453 2'-O-methyltransferase rep ORF1ab polyprotein 1a-1b protein protein cvemc Middle East respiratory syndrome-related coronavirus (isolate United Kingdom/H123990006/2012) (Betacoronavirus England 1) Betacoronavirus England 1 Human coronavirus EMC ASADWKPGHAMPSLFKVQNVNLERCELANYKQSIPMPRGVHMNIAKYMQLCQYLNTCTLAVPANMRVIHFGAGSDKGIAPGTSVLRQWLPTDAIIIDNDLNEFVSDADITLFGDCVTVRVGQQVDLVISDMYDPTTKNVTGSNESKALFFTYLCNLINNNLALGGSVAIKITEHSWSVELYELMGKFAWWTVFCTNANASSSEGFLLGINYLGTIKENIDGGAMHANYIFWRNSTPMNLSTYSLFDLSKFQLKLKGTPVLQLKESQINELVISLLSQGKLLIRDNDTLSVSTDVLVNTYRKLR 6776 7078 3C6BDE237B9463F9 p0c6x7-pro_0000037317 Non-structural protein 10 rep ORF1ab polyprotein 1a-1b protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus AGNATEVPANSTVLSFCAFAVDPAKAYKDYLASGGQPITNCVKMLCTHTGTGQAITVTPEANMDQESFGGASCCLYCRCHIDHPNPKGFCDLKGKYVQIPTTCANDPVGFTLRNTVCTVCGMWKGYGCSCDQLREPLMQ 4231 4369 1F48DD3A6874826C q98vg9-pro_5000140223 Putative 2'-O-methyl transferase rep ORF1ab polyprotein 1a-1b protein protein FCoV Feline coronavirus (strain FIPV WSU-79/1146) SAEWNPGYSMPTLYKIQRMCLERCNLYNYGAQVRLPDGITTNVVKYTQLCQYLNTTTVCVPHKMRVLHLGAAGASGVAPGSTVLRRWLPDDAILVDNDLRDYVSDADFSVTGDCTSLYIEDKFDLLISDLYDGSTKSIDGENTSKDGFFTYINGFIKEKLSLGGSAAIKITEFSWNKDLYELIQRFEYWTVFCTSVNTSSSEGFLIGINYLGPYCDKAIVDGNIMHANYIFWRNSTIMALSHNSVLDTPKFKCRCNNALIVNLKEKELNEMVVGLLRKGKLLIRNNGKLLNFGNHLVNVP 6410 6709 C3042E7AC26649C3 p0c6y0-pro_0000037368 2'-O-methyl transferase rep ORF1ab polyprotein 1a-1b protein protein MHV-JHM Murine coronavirus (strain JHM) Murine hepatitis virus AAADWKPGYVMPVLYKYLESPMERVNLWNYGKPITLPTGCMMNVAKYTQLCQYLSTTTLAVPANMRVLHLGAGSDKGVAPGSAVLRQWLPSGSILVDNDMNPFVSDSVASYYGNCITLPFDCQWDLIISDMYDPLTKNIGEYNVSKDGFFTYLCHLIRDKLALGGSVAIKITEFSWNAELYSLMGKFAFWTIFCTNVNASSSEGFLIGINWLNRTRNEIDGKTMHANYLFWRNSTMWNGGAYSLFDMTKFPLKAAGTAVVSLKPDQINDLVLSLIEKGKLLVRDTRKEVFVGDSLVNVK 6882 7180 EE81BAB7014C0235 p0c6y1-pro_0000037417 2'-O-methyl transferase rep ORF1ab polyprotein 1a-1b protein protein ibvb ibvb IBV SAWTCGYNMPELYKVQNCVMEPCNIPNYGVGITLPSGILMNVAKYTQLCQYLSKTTICVPHNMRVMHFGAGSDKGVAPGSTVLKQWLPEGTLLVDNDIVDYVSDAHVSVLSDCNKYNTEHKFDLVISDMYTDNDSKRKHEGVIANNGNDDVFIYLSSFLRNNLALGGSFAVKVTETSWHEVLYDIAQDCAWWTMFCTAVNASSSEAFLIGVNYLGASEKVKVSGKTLHANYIFWRNCNYLQTSAYSIFDVAKFDLRLKATPVVNLKTEQKTDLVFNLIKCGKLLVRDVGNTSFTSDSFVCTM 6328 6629 15ECCB26B9D5423D paps-pppac20_rna_substrate-2 2 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro 8 enzyme enzyme neutral component neutral component in vitro In vitro methylation methylation reaction methylation 1B-G, 3, 4 Positive control experiment; no information is given with respect to purification, expression or construct details for VP39. paps-pppac20_rna_substrate-3 2 3 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 4 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0C6Y5-PRO_0000037395:p.Lys93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A P0C6Y5-PRO_0000037395:p.Phe96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1B, 2, 3C, 4A, 5A TGEV (Porcine transmissible gastroenteritis coronavirus ) nsp10+16 proteins. paps-pppac20_rna_substrate-1 2 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain in vitro In vitro 6 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range Q98VG9-PRO_5000140217:p.Lys93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A Q98VG9-PRO_5000140217:p.Phe96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) methylation methylation reaction methylation 1C, number 1, 2, 3C, 4B, 5A FCoV (feline coronavirus) nsp10+16 proteins. pppac20_rna_substrate-rep-1 2 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain in vitro In vitro methylation methylation reaction methylation 1C, number 2 FCoV (feline coronavirus) 16 protein. FCoV nsp16 alone showed a low but detectable MTase activity. pppac20_rna_substrate-rep-2 2 15 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0C6Y0-PRO_0000037363:p.Arg93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R A P0C6Y0-PRO_0000037363:p.Phe96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1D, 3D, 4C, 5A MHV (mouse hepatitis virus) nsp10+16 proteins. pppac20_rna_substrate-rep-3 2 10 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 13 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0C6X7-PRO_0000037317:p.Tyr96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain Y A P0C6X7-PRO_0000037317:p.Lys93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1E, 2, 3D, 4D, 5A SARS-CoV nsp10+16 proteins. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-4 2 12 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 9 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range K9N7C7-PRO_0000422448:p.Lys93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A K9N7C7-PRO_0000422448:p.Phe96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1F, 3D, 4E, 5A MERS-CoV nsp10+16 proteins. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-5 2 16 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0C6Y1-PRO_0000037412:p.Lys98Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A P0C6Y1-PRO_0000037412:p.Phe101Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1G, 2, 4F, 5A IBV (infectious bronchitis virus) nsp10+16 proteins. Identity of "IBV" confirmed from Ref.28 (PMID: 22022266) , Supp. figure 2 sequence. The nsp16 of IBV (Gammacoronavirus) was activated and stimulated y only by its cognate nsp10. Current Uniprot P0C6Y1 sequence numbering for IBV nsp10 differs from that given in figure 3A. pppac20_rna_substrate-rep-6 1 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1C, number 2 FCoV (feline coronavirus) 16 protein. FCoV nsp16 alone generated a small fraction of products that comigrated with m7G*pppAm structure. showed a low but detectable MTase activity. pppac20_rna_substrate-rep-7 1 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro 6 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) methylation methylation reaction methylation 1C, number 1 FCoV (feline coronavirus) nsp10+16 proteins. pppac20_rna_substrate-rep-8 1 10 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 13 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain resulting-cleavage resulting-cleavage in vitro In vitro methylation methylation reaction methylation 1E SARS-CoV nsp10+16 proteins. paps-pppac20_rna_substrate-4 1 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro 8 enzyme enzyme neutral component neutral component in vitro In vitro methylation methylation reaction methylation 1H Positive control experiment; no information is given with respect to purification, expression or construct details for VP39. pppac20_rna_substrate-rep-9 1 3 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 4 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain in vitro In vitro methylation methylation reaction methylation 1B TGEV (Porcine transmissible gastroenteritis coronavirus ) nsp10+16 proteins. pppac20_rna_substrate-rep-10 1 15 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1D MHV (mouse hepatitis virus) nsp10+16 proteins. pppac20_rna_substrate-rep-11 1 12 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 9 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm atp 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1F MERS-CoV nsp10+16 proteins. pppac20_rna_substrate-rep-12 1 16 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 1G IBV (infectious bronchitis virus) nsp10+16 proteins. Identity of "IBV" confirmed from Ref.28 (PMID: 22022266) , Supp. figure 2 sequence. pppac20_rna_substrate-rep-13 2 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain in vitro In vitro 4 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) methylation methylation reaction methylation 3C, 4B FCoV (feline coronavirus) nsp16 protein and TGEV (Porcine transmissible gastroenteritis coronavirus ) nsp10 protein. FCoV nsp16 could be stimulated to a similar level either by cognate nsp10 or noncognate nsp10s. pppac20_rna_substrate-rep-14 2 3 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 6 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 3C, 4A TGEV (Porcine transmissible gastroenteritis coronavirus ) nsp16 protein and FCov nsp10 protein. nsp16 of TGEV could be fully stimulated by the nsp10 of Alphacoronaviruses (FCov). pppac20_rna_substrate-rep-15 2 15 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 13 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 3D, 4C MHV (mouse hepatitis virus) 16 protein and SarsCoV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-16 2 15 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 9 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 3D, 4C MHV (mouse hepatitis virus) 16 protein and MersCoV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-17 2 12 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 13 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 3D, 4E MERS-CoV 16 protein and SarsCoV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-18 2 12 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 3D, 4E MERS-CoV 16 protein and MHV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-19 2 3 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4A Weak interaction;TGEV (Porcine transmissible gastroenteritis coronavirus) nsp16 protein and MHV nsp10 protein. nsp16 of TGEV could be fully stimulated by the nsp10 of Alphacoronavirus but less efficiently activated when using the nsp10 of Betacoronavirus and Gammacoronavirus. pppac20_rna_substrate-rep-20 2 3 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 13 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4A Weak interaction;TGEV (Porcine transmissible gastroenteritis coronavirus) nsp16 protein and SarsCoV nsp10 protein. nsp16 of TGEV could be fully stimulated by the nsp10 of Alphacoronavirus but less efficiently activated when using the nsp10 of Betacoronavirus and Gammacoronavirus. pppac20_rna_substrate-rep-21 2 3 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4A Weak interaction;TGEV (Porcine transmissible gastroenteritis coronavirus) nsp16 protein and IBV nsp10 protein. nsp16 of TGEV could be fully stimulated by the nsp10 of Alphacoronavirus but less efficiently activated when using the nsp10 of Betacoronavirus and Gammacoronavirus. pppac20_rna_substrate-rep-22 2 3 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 9 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4A Weak interaction;TGEV (Porcine transmissible gastroenteritis coronavirus) nsp16 protein and Mers-CoV nsp10 protein. nsp16 of TGEV could be fully stimulated by the nsp10 of Alphacoronavirus but less efficiently activated when using the nsp10 of Betacoronavirus and Gammacoronavirus. pppac20_rna_substrate-rep-23 2 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain in vitro In vitro 5 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) methylation methylation reaction methylation 4B FCoV (feline coronavirus) nsp16 protein and MHV nsp10 protein. FCoV nsp16 could be stimulated to a similar level either by cognate nsp10 or noncognate nsp10s. pppac20_rna_substrate-rep-24 2 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain in vitro In vitro 13 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) methylation methylation reaction methylation 4B FCoV (feline coronavirus) nsp16 protein and SARS-CoV nsp10 protein. FCoV nsp16 could be stimulated to a similar level either by cognate nsp10 or noncognate nsp10s. pppac20_rna_substrate-rep-25 2 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain in vitro In vitro 9 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) methylation methylation reaction methylation 4B FCoV (feline coronavirus) nsp16 protein and MERS-CoV nsp10 protein. FCoV nsp16 could be stimulated to a similar level either by cognate nsp10 or noncognate nsp10s. pppac20_rna_substrate-rep-26 2 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain in vitro In vitro 11 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) methylation methylation reaction methylation 4B FCoV (feline coronavirus) nsp16 protein and IBV nsp10 protein. FCoV nsp16 could be stimulated to a similar level either by cognate nsp10 or noncognate nsp10s. pppac20_rna_substrate-rep-27 2 15 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 4 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4C MHV (mouse hepatitis virus) 16 protein and TGEV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-28 2 15 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 6 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4C MHV (mouse hepatitis virus) 16 protein and FCoV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-29 2 15 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4C MHV (mouse hepatitis virus) 16 protein and IBV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-30 2 10 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 3D, 4D SARS-CoV 16 proteins and MHV nsp10. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-31 2 10 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 9 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 3D, 4D SARS-CoV 16 proteins and MersCoV nsp10. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-32 2 10 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 4 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4D SARS-CoV 16 proteins and TGEV nsp10. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-33 2 10 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 6 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4D SARS-CoV 16 proteins and FCoV nsp10. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-34 2 10 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4D SARS-CoV 16 proteins and IBV nsp10. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-35 2 12 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 4 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4E MERS-CoV 16 protein and TGEV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-36 2 12 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 6 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4E MERS-CoV 16 protein and FCoV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-37 2 12 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 4E MERS-CoV 16 protein and IBV nsp10 protein. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-38 2 5 inhibitor inhibitor neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal Tyr ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res certain certain sequence position certain certain certain sequence position certain C-terminal Phe amidated residue amidated residue certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 6 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 14 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain in vitro In vitro methylation methylation reaction methylation 5B, number 1 FCoV (feline coronavirus) nsp10+16 proteins. pppac20_rna_substrate-rep-39 2 5 inhibitor inhibitor neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal Tyr ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res certain certain sequence position certain certain certain sequence position certain C-terminal Phe amidated residue amidated residue certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 3 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 4 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0C6Y5-PRO_0000037395:p.Lys93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A P0C6Y5-PRO_0000037395:p.Phe96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 5B number 2 TGEV (Porcine transmissible gastroenteritis coronavirus ) nsp10+16 proteins. pppac20_rna_substrate-rep-40 2 5 inhibitor inhibitor neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal Tyr ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res certain certain sequence position certain certain certain sequence position certain C-terminal Phe amidated residue amidated residue certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 15 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 5 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0C6Y0-PRO_0000037363:p.Arg93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R A P0C6Y0-PRO_0000037363:p.Phe96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 5B, number 3 MHV (mouse hepatitis virus) nsp10+16 proteins. pppac20_rna_substrate-rep-41 2 5 inhibitor inhibitor neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal Tyr ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res certain certain sequence position certain certain certain sequence position certain C-terminal Phe amidated residue amidated residue certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 10 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 13 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0C6X7-PRO_0000037317:p.Tyr96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain Y A P0C6X7-PRO_0000037317:p.Lys93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 5B, number 4 SARS-CoV nsp10+16 proteins. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-42 2 5 inhibitor inhibitor neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal Tyr ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res certain certain sequence position certain certain certain sequence position certain C-terminal Phe amidated residue amidated residue certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 12 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 9 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range K9N7C7-PRO_0000422448:p.Lys93Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A K9N7C7-PRO_0000422448:p.Phe96Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 5B, number 5 MERS-CoV nsp10+16 proteins. nsp16s of Betacoronavirus, including MHV, SARS-CoV , and MERS-CoV, could be fully activated and stimulated with the help of nsp10s of all selected coronaviruses from different genera. pppac20_rna_substrate-rep-43 2 5 inhibitor inhibitor neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal Tyr ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res certain certain sequence position certain certain certain sequence position certain C-terminal Phe amidated residue amidated residue certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 16 enzyme enzyme neutral component neutral component purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 11 stimulator stimulator ancillary ancillary purified purified n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range P0C6Y1-PRO_0000037412:p.Lys98Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain K A P0C6Y1-PRO_0000037412:p.Phe101Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain F A ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 5'_methyl_N7_Guanosine rna chemical modification RNA chemical modification RNA chemical modification post-transcriptional modification rna chemical modification certain certain sequence position certain certain certain sequence position certain 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm in vitro In vitro methylation methylation reaction methylation 5B IBV (infectious bronchitis virus) nsp10+16 proteins. Identity of "IBV" confirmed from Ref.28 (PMID: 22022266) , Supp. figure 2 sequence. The nsp16 of IBV (Gammacoronavirus) was activated and stimulated y only by its cognate nsp10. Current Uniprot P0C6Y1 sequence numbering for IBV nsp10 differs from that given in figure 3A.