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/ NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. Nucleic acids research 2021 imex curation Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus artsimovitch.1@osu.edu Only protein-protein interactions imex curation in vitro In vitro cleavage cleavage assay cleavage tag fluorescence tag visualisation by fluorescence tag fluorescence Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Pyrophosphorolysis assay:the reverse of the polymerisation reaction in which pyrophosphate reacts with the 3′ nucleotidemonophosphate (NMP or dNMP), which is removed from the oligonucleotide to release the corresponding triphosphate. artsimovitch.1@osu.edu Accepted 2021-NOV-10 AT 15:53 GMT AT 15:53 GMT by LUANA Nucleic acids research 2021 Only protein-protein interactions imex curation imex curation NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. Nucleic acids research 2021 imex curation Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus artsimovitch.1@osu.edu Only protein-protein interactions imex curation in vitro In vitro gtpase assay gtpase assay GTPase gtp hydrolisis gtp hydrolisis GTPase predetermined predetermined participant predetermined Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Hydrolysis of radiolabeled GTP was observed using thin layer chromatography. artsimovitch.1@osu.edu Accepted 2021-NOV-10 AT 15:53 GMT AT 15:53 GMT by LUANA Nucleic acids research 2021 Only protein-protein interactions imex curation imex curation NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. Nucleic acids research 2021 imex curation Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus artsimovitch.1@osu.edu Only protein-protein interactions imex curation in vitro In vitro cleavage cleavage assay cleavage predetermined predetermined participant predetermined Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Pyrophosphorolysis assay:the reverse of the polymerisation reaction in which pyrophosphate reacts with the 3′ nucleotidemonophosphate (NMP or dNMP), which is removed from the oligonucleotide to release the corresponding triphosphate. The formation of radiolabeled GTP/GDP was observed using thin layer chromatography, after the addition of PP i. artsimovitch.1@osu.edu Accepted 2021-NOV-10 AT 15:53 GMT AT 15:53 GMT by LUANA Nucleic acids research 2021 Only protein-protein interactions imex curation imex curation NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. Nucleic acids research 2021 imex curation Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus artsimovitch.1@osu.edu Only protein-protein interactions imex curation in vitro In vitro enzymatic study enzymatic study experimental particp experimental participant identification experimental particp Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Nucleotidyl-monophosphate transferase activity-NMPylation;protein guanylylation. artsimovitch.1@osu.edu Accepted 2021-NOV-10 AT 15:53 GMT AT 15:53 GMT by LUANA Nucleic acids research 2021 Only protein-protein interactions imex curation imex curation NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. Nucleic acids research 2021 imex curation Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus artsimovitch.1@osu.edu Only protein-protein interactions imex curation in vitro In vitro enzymatic study enzymatic study weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins de-NMPylation:Reverse guanylyltransferase activity;pre-GMPylated nsp9 was incubated with WT or mutant nsp12. PPi was then added. As a result GMP is removed from Nsp9. artsimovitch.1@osu.edu Accepted 2021-NOV-10 AT 15:53 GMT AT 15:53 GMT by LUANA Nucleic acids research 2021 Only protein-protein interactions imex curation imex curation NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. NMPylation and de-NMPylation of SARS-CoV-2 nsp9 by the NiRAN domain. Nucleic acids research 2021 imex curation Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus artsimovitch.1@osu.edu Only protein-protein interactions imex curation in vitro In vitro rna rna pol assay rna directed rna polymerase assay rna rna pol assay tag fluorescence tag visualisation by fluorescence tag fluorescence Wang B., Svetlov D., Artsimovitch I. Virus - Publications including interactions involving viral proteins artsimovitch.1@osu.edu Accepted 2021-NOV-10 AT 15:53 GMT AT 15:53 GMT by LUANA Nucleic acids research 2021 Only protein-protein interactions imex curation imex curation Mock publication and experiment for abstract interactions that are not interaction evidences. Mock publication and experiment for abstract interactions that are not interaction evidences. unspecified method nucleoside triphosphate nucleoside triphosphate NTP Nucleoside triphosphate nucleoside triphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) p0dtd1-pro_0000449629 RNA-directed RNA polymerase nsp12 rep ORF1ab polyprotein 1a-1b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV SADAQSFLNRVCGVSAARLTPCGTGTSTDVVYRAFDIYNDKVAGFAKFLKTNCCRFQEKDEDDNLIDSYFVVKRHTFSNYQHEETIYNLLKDCPAVAKHDFFKFRIDGDMVPHISRQRLTKYTMADLVYALRHFDEGNCDTLKEILVTYNCCDDDYFNKKDWYDFVENPDILRVYANLGERVRQALLKTVQFCDAMRNAGIVGVLTLDNQDLNGNWYDFGDFIQTTPGSGVPVVDSYYSLLMPILTLTRALTAESHVDTDLTKPYIKWDLLKYDFTEERLKLFDRYFKYWDQTYHPNCVNCLDDRCILHCANFNVLFSTVFPPTSFGPLVRKIFVDGVPFVVSTGYHFRELGVVHNQDVNLHSSRLSFKELLVYAADPAMHAASGNLLLDKRTTCFSVAALTNNVAFQTVKPGNFNKDFYDFAVSKGFFKEGSSVELKHFFFAQDGNAAISDYDYYRYNLPTMCDIRQLLFVVEVVDKYFDCYDGGCINANQVIVNNLDKSAGFPFNKWGKARLYYDSMSYEDQDALFAYTKRNVIPTITQMNLKYAISAKNRARTVAGVSICSTMTNRQFHQKLLKSIAATRGATVVIGTSKFYGGWHNMLKTVYSDVENPHLMGWDYPKCDRAMPNMLRIMASLVLARKHTTCCSLSHRFYRLANECAQVLSEMVMCGGSLYVKPGGTSSGDATTAYANSVFNICQAVTANVNALLSTDGNKIADKYVRNLQHRLYECLYRNRDVDTDFVNEFYAYLRKHFSMMILSDDAVVCFNSTYASQGLVASIKNFKSVLYYQNNVFMSEAKCWTETDLTKGPHEFCSQHTMLVKQGDDYVYLPYPDPSRILGAGCFVDDIVKTDGTLMIERFVSLAIDAYPLTKHPNQEYADVFHLYLQYIRKLHDELTGHMLDMYSVMLTNDNTSRYWEPEFYEAMYTPHTVLQ 4393 5324 gtp guanosine triphosphate - GTP 5'-GTP GTP Guanosine 5'-triphosphate guanosine 5'-triphosphoric acid guanosine triphosphate GUANOSINE-5'-TRIPHOSPHATE H4gtp guanosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 p0dtd1-pro_0000449626 Non-structural protein 8 rep ORF1ab polyprotein 1a-1b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV AIASEFSSLPSYAAFATAQEAYEQAVANGDSEVVLKKLKKSLNVAKSEFDRDAAMQRKLEKMADQAMTQMYKQARSEDKRAKVTSAMQTMLFTMLRKLDNDALNNIINNARDGCVPLNIIPLTTAAKLMVVIPDYNTYKNTCDGTTFTYASALWEIQQVVDADSKIVQLSEISMDNSPNLAWPLIVTALRANSAVKLQ 3943 4140 sodium diphosphate sodium diphosphate Diphosphoric acid, tetrasodium salt Natrium pyrophosphat Pyrophosphoric acid tetrasodium salt Sodium pyrophosphate Tetrasodium pyrophosphate TSPP sodium diphosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) p0dtd1-pro_0000449627 Non-structural protein 9 rep ORF1ab polyprotein 1a-1b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV NNELSPVALRQMSCAAGTTQTACTDDNALAYYNTTKGGRFVLALLSDLQDLKWARFPKSDGTGTIYTELEPPCRFVTDTPKGPKVKYLYFIKGLNNLNRGMVLGSLAATVRLQ 4141 4253 p0dtd1-pro_0000449625 Non-structural protein 7 rep ORF1ab polyprotein 1a-1b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV SKMSDVKCTSVVLLSVLQQLRVESSSKLWAQCVQLHNDILLAKDTTEAFEKMVSLLSVLLSMQGAVDINKLCEEMLDNRATLQ 3860 3942 gmp GMP 5'-GMP GMP Guanosine 5'-monophosphate Guanosine 5'-phosphate Guanosine monophosphate guanosine-5'-monophosphate Guanylic acid pG 5'-guanylic acid [(2R,3S,4R,5R)-5-(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl dihydrogen phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) RQFCJASXJCIDSX-UUOKFMHZSA-N RQFCJASXJCIDSX-UUOKFMHZSA-N sodium pyrophosphate decahydrate Sodium pyrophosphate decahydrate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) rna scaffold rna ribonucleic acid RNA rna chemical synthesis Chemical synthesis (Chemical synthesis) UUUUCAUGCUACGCGUAGUUUUCUACGCG rep-rep-266 4 10 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp enzyme enzyme neutral component neutral component purified purified P0DTD1-PRO_0000449629:p.Asp760Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain D A Tue Oct 05 18:58:30 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp760Ala' P0DTD1-PRO_0000449629:p.Asp218Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A Tue Oct 05 18:53:24 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p. Asp218Ala' P0DTD1-PRO_0000449629:p.Lys50Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain K A Tue Oct 05 19:20:44 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys50Ala' P0DTD1-PRO_0000449629:p.Arg116Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R A Tue Oct 05 18:55:13 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg116Ala' P0DTD1-PRO_0000449629:p.Tyr129Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain Y A Tue Oct 05 19:01:12 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr129Ala' ecoli Escherichia coli (strain K12) 14 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra sequence tag sequence tag identification sequence tag experimental particp experimental participant identification experimental particp enzyme target enzyme target substrate substrate neutral component neutral component purified purified p.Asn1 guanylated residue guanylated residue 5'phos Guanosyl Phosphoguanosine phospho-guanosine certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm P0DTD1-PRO_0000449627:p.Asn1delinsGlySerAsn mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain N GSN Tue Oct 05 19:38:20 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.1delinsGlySerAsn' ecoli Escherichia coli (strain K12) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction 1 C/D, 3C, Supp. figure 5, 3 the 3 aas at the N-terminus will almost certainly throw a wobbly for the mutation update as there is no way of showing an N-terminal mutation in HGVS. (see my email at EBI for August 2020 from JdD.nl for tRNA similar situation- 5' extension) Preincubation of nsp12 with a three-fold molar excess of GSNnsp9 only slightly inhibited modification of Nnsp9. The N-terminus of nsp9 is modified by GMP. rep-rep-267 4 13 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 10 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 14 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra experimental particp experimental participant identification experimental particp enzyme target enzyme target substrate substrate neutral component neutral component purified purified p.Asn1 guanylated residue guanylated residue 5'phos Guanosyl Phosphoguanosine phospho-guanosine certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ecoli Escherichia coli (strain K12) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction 2 B/C, 3B,4B As well as PPi, several nucleotides or their analogues acted as competitors or inhibitors of the interaction, notably; NTPs, GDP, GMP, ITP, and GMPCPP. The bisphosphonates Foscarnet (Fos- CHEBI:127780) and Risedronate (Ris-CHEBI:8868) inhibit nsp9 NMPylation, although less efficiently than PPi. ppGpp and RTP (Remdesivir triphosphate) were poor competitors/ inhibitors. maybe strictly speaking, interactions with these competitors/ inhibitors should be added - but as they do not know which type (competitive inhibition of GTP binding (e.g. by GDP or GMP) or to nsp9 modification) they are it did not seem too useful info. nsp7_nsp8_nsp12_sars2-rna scaffold-1 1 18 enzyme target enzyme target substrate substrate neutral component neutral component 5' prime cy5 label cy5 label certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 38 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 17 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage 3A LHS I do not know quite what they refer to in the figure legend text:- "Bottom: SARS-CoV-2 RdRp transcription complex assembled on the 5′ Cy5.5-labeled hairpin, which comprises both the template and the product RNAs, is completely resistant to PPi even in the absence of NTPs. " - there is NO figure at the bottom?? nsp7_nsp8_nsp12_sars2-rna scaffold-2 1 8 inhibitor inhibitor neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 18 enzyme target enzyme target substrate substrate neutral component neutral component 5' prime cy5 label cy5 label certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 38 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 17 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage 3A LHS, Supp. figure 6 In the presence of non-cognate NTPs, SARS-CoV-2 RdRp did not cleave nascent RNA. nsp7_nsp8_nsp12_sars2-nucleoside triphosphate-1 6 18 enzyme target enzyme target substrate substrate neutral component neutral component 5' prime cy5 label cy5 label certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 38 enzyme enzyme neutral component neutral component purified purified P0DTD1-PRO_0000449629:p.Asp760Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A 51 Tue Oct 05 19:28:54 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp760Ala' P0DTD1-PRO_0000449629:p.Asp218Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain D A 51 Tue Oct 05 18:31:22 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp218Ala' P0DTD1-PRO_0000449629:p.Arg116Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain R A 51 Tue Oct 05 19:01:28 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg116Ala' P0DTD1-PRO_0000449629:p.Lys50Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain K A 51 Tue Oct 05 19:09:23 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys50Ala' P0DTD1-PRO_0000449629:p.Tyr129Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain Y A 51 Tue Oct 05 19:03:11 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr129Ala' ecoli Escherichia coli (strain K12) 8 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage 3A RHS, Supp. figure 1 No cleavage was seen in the presence of PPi (-CHEBI:177382). rep-rep-268 5 13 enzyme regulator enzyme regulator neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 10 predetermined predetermined participant predetermined weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra enzyme enzyme neutral component neutral component purified purified P0DTD1-PRO_0000449629:p.Asp760Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain D A Tue Oct 05 19:22:18 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp760Ala' P0DTD1-PRO_0000449629:p.Asp218Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain D A Tue Oct 05 18:38:44 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp218Ala' ecoli Escherichia coli (strain K12) 14 enzyme target enzyme target substrate substrate neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 16 enzyme target enzyme target substrate substrate neutral component neutral component region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 63 66 cleavage cleavage reaction cleavage 3C, Supp. figure 7 GMP is removed from Nsp9. rep-rep-269 4 38 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 14 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra sequence tag sequence tag identification sequence tag experimental particp experimental participant identification experimental particp enzyme target enzyme target substrate substrate neutral component neutral component purified purified p.Asn1 guanylated residue guanylated residue 5'phos Guanosyl Phosphoguanosine phospho-guanosine certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm P0DTD1-PRO_0000449627:p.Asn1delinsGlySerAsn mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain N GSN Tue Oct 05 18:37:39 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.1delinsGlySerAsn' ecoli Escherichia coli (strain K12) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction Supp. figure 3B The N-terminus of nsp9 is modified by GMP. nsp7_nsp8_nsp12_sars2-rep-1 4 18 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 38 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 14 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra sequence tag sequence tag identification sequence tag experimental particp experimental participant identification experimental particp enzyme target enzyme target substrate substrate neutral component neutral component purified purified p.Asn1 guanylated residue guanylated residue 5'phos Guanosyl Phosphoguanosine phospho-guanosine certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm P0DTD1-PRO_0000449627:p.Asn1delinsGlySerAsn mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain N GSN Tue Oct 05 19:33:15 BST 2021 This feature has been corrected as a result of our quality control procedures. The original label was 'p.1delinsGlySerAsn' ecoli Escherichia coli (strain K12) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction Supp. figure 3B The N-terminus of nsp9 is modified by GMP. nsp7_nsp8_nsp12_sars2-nucleoside triphosphate-2 6 18 enzyme target enzyme target substrate substrate neutral component neutral component 5' prime cy5 label cy5 label certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 38 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 9 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) cleavage cleavage reaction cleavage Supp. figure 6 RdRp misincorporates some AMP to generate a 40 residue product. gmp-rep-1 5 13 enzyme regulator enzyme regulator neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 38 predetermined predetermined participant predetermined weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 14 enzyme target enzyme target substrate substrate neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 16 enzyme target enzyme target substrate substrate neutral component neutral component region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 90 87 cleavage cleavage reaction cleavage Supp. figure 7 GMP is removed from Nsp9. gmp-rep-2 3 13 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 10 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 14 enzyme target enzyme target substrate substrate neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 16 enzyme target enzyme target substrate substrate neutral component neutral component region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 95 98 cleavage cleavage reaction cleavage Supp. figure 8, Lane 6 gmp-rep-3 2 10 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 11 enzyme target enzyme target substrate substrate neutral component neutral component region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) gtpase reaction gtpase reaction Supp. figure 8, Lane 7 nsp12 shows unusual intrinsic nucleotide hydrolytic activity. nsp7-nsp8-nsp12_sars2 SARS-CoV-2 polymerase complex NSP7:2xNSP8:NSP12 replication-transcription complex replication and transcription complex RTC central RTC C-RTC holo-RdRp complex holo-RNA-dependent RNA polymerase complex 12 enzyme enzyme nsp7_nsp8 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 enzyme enzyme nsp7_nsp8 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 enzyme enzyme nsp8_nsp12 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 15 unspecified role unspecified role nsp8_nsp12 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 109 104 107 104 109 105 105 107 physical association physical association SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV stable complex stable complex physical interaction evidence used in manual assertion physical interaction evidence used in manual assertion IPI inferred from physical interaction RNA-directed 5'-3' RNA polymerase of the SARS-CoV-2 coronavirus which consists of the main polymerase protein NSP12 and a stoichiometric variant of the primase complex (CPX-5690). Extends partially double-stranded RNA templates and is probably also required for transcription initiation, though the mechanism for this has yet to be determined. Complex formation enhances dsRNA binding of the individual protomers. Heterotetramer Severe acute respiratory syndrome (SARS) [EFO:0000694]: A viral respiratory infection caused by the SARS coronavirus. It is transmitted through close person-to-person contact. It is manifested with high fever, headache, dry cough and myalgias. It may progress to pneumonia and cause death. Primase complex without polymerase has a different stoichiometry (8:8). NSP12 possesses conserved motifs involved in template binding, NTP binding, and polymerization that assume a structure resembling a cupped “right hand”. The NSP7-NSP8 heterodimer binds to the index finger loop and may facilitates the interaction of NSP12 with additional components of the RNA synthesis machinery. The second NSP8 molecule binds close to the RNA template-binding channel. Accepted 2020-APR-20 by ORCHARD coronavirus