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/ http://www.ebi.ac.uk/ ke-2012-2 in vitro In vitro methyltransferase as methyltransferase assay methyltransferase as autoradiography autoradiography Ke M., Chen Y., Wu A., Sun Y., Su C., Wu H., Jin X., Tao J., Wang Y., Ma X., Pan JA., Guo D. Virus - Publications including interactions involving viral proteins Reaction mixtures were spotted onto TLC plates. Radiolabelling was determined using a PhosphorImager. dguo@whu.edu.cn Virus research 2012 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus ke-2012-3 in vitro In vitro crosslink cross-linking study crosslink autoradiography autoradiography Ke M., Chen Y., Wu A., Sun Y., Su C., Wu H., Jin X., Tao J., Wang Y., Ma X., Pan JA., Guo D. Virus - Publications including interactions involving viral proteins SAM binding assay:protein mixtures incubated with S-adenosyl [methyl-3H] methionine were irradiated with UV light to cross-link SAM to its binding site. Samples were separated by SDS-PAGE and and analyzed by autoradiography. dguo@whu.edu.cn Virus research 2012 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus ke-2012-4 in vitro In vitro methyltransferase as methyltransferase assay methyltransferase as tag visualisation tag visualisation Ke M., Chen Y., Wu A., Sun Y., Su C., Wu H., Jin X., Tao J., Wang Y., Ma X., Pan JA., Guo D. Virus - Publications including interactions involving viral proteins Radiolabeled S-adenosyl [methyl-3H] methionine, purified SARS-CoV nsp16/nsp10 complex, +/- short peptides, was mixed with m7GpppA-RNA substrate. The 3H-labeled product was isolated in small DEAE-Sephadex columns and quantitated by liquid scintillation counting. dguo@whu.edu.cn Virus research 2012 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus ke-2012-1 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Ke M., Chen Y., Wu A., Sun Y., Su C., Wu H., Jin X., Tao J., Wang Y., Ma X., Pan JA., Guo D. Virus - Publications including interactions involving viral proteins dguo@whu.edu.cn Virus research 2012 Only protein-protein interactions imex curation imex curation Coronavirus - Interactions investigated in the context of Coronavirus 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 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 sars_rna_substrate rna ribonucleic acid RNA rna synthetic construct synthetic construct AGAUUAGGUUUUUACCUACCCAGGAAAAGCCAACCAACCUCGAUCUCUUGUAGAUCUGUUCUCUAAACGAACUUUAAAAUCUGUGUAGCUGUCGCUCGGCUGCAUGCCUAGUGCACCUACGCAGUAUAAACAAUAAUAAAUUUUACUGUCGUUGACAAGAAACGAGUAACUCGUCCCUCUUCUGCAGACUGCUUACGGUUUCGUCCGUGUUGCAGUCGAUCAUCAGCAUACCUAGGUUUCGUCCGGGUGUGACCGAAAG 5′-terminal 259 nucleotides of the SARS-CoV genome, strain WHU, GenBank: AY394850.2, with first U mutated to a G (pmid:19208801) p0c6x7-pro_0000037320 Proofreading exoribonuclease nsp14 rep ORF1ab polyprotein 1a-1b protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus AENVTGLFKDCSKIITGLHPTQAPTHLSVDIKFKTEGLCVDIPGIPKDMTYRRLISMMGFKMNYQVNGYPNMFITREEAIRHVRAWIGFDVEGCHATRDAVGTNLPLQLGFSTGVNLVAVPTGYVDTENNTEFTRVNAKPPPGDQFKHLIPLMYKGLPWNVVRIKIVQMLSDTLKGLSDRVVFVLWAHGFELTSMKYFVKIGPERTCCLCDKRATCFSTSSDTYACWNHSVGFDYVYNPFMIDVQQWGFTGNLQSNHDQHCQVHGNAHVASCDAIMTRCLAVHECFVKRVDWSVEYPIIGDELRVNSACRKVQHMVVKSALLADKFPVLHDIGNPKAIKCVPQAEVEWKFYDAQPCSDKAYKIEELFYSYATHHDKFTDGVCLFWNCNVDRYPANAIVCRFDTRVLSNLNLPGCDGGSLYVNKHAFHTPAFDKSAFTNLKQLPFFYYSDSPCESHGKQVVSDIDYVPLKSATCITRCNLGGAVCRHHANEYRQYLDAYNMMISAGFSLWIYKQFDTYNLWNTFTRLQ 5903 6429 s-adenosyl-l-methionine S adenosyl Methionine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) nsp10_nsp16_cvhsa-sars_rna_substrate-1 3725355 N/A 3725359 ancillary ancillary neutral component neutral component purified purified region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 3725360 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 false ecoli Escherichia coli (strain K12) N/A 3725361 enzyme target enzyme target substrate substrate neutral component neutral component ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain false 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain false 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 false in vitro In vitro methylation methylation reaction methylation false false false 2C, Lanes 12,15,16. rep-rep-91 3725355 N/A 3725362 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 false ecoli Escherichia coli (strain K12) N/A 3725360 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 false ecoli Escherichia coli (strain K12) N/A 3725359 ancillary ancillary neutral component neutral component purified purified region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 3725361 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 false 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain false ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain false 5'_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 false in vitro In vitro methylation methylation reaction methylation false false false 2C, Lanes 5,6,2. A two-staged reaction: overall reaction = nsp10/nsp14/nsp16 methylates G*pppA-RNA to m7G*pppAm-RNA (Fig. 2C, lane 2): nsp14 first methylates G*pppA-RNA at N7 position of guanine (m7G) to generate m7GpppA-RNA, (Fig. 2C, lanes 1 and 7), followed by the methylation of m7G*pppA-RNA at the 2′-O-position of adenine (Am) to generate m7G*pppAm-RNA. rep-rep-92 3725355 N/A 3725362 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 false ecoli Escherichia coli (strain K12) N/A 3725361 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 false ATP 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain false 5'_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 false in vitro In vitro methylation methylation reaction methylation false false false 2C, Lanes 1,7. Reaction: nsp14 methylates G*pppA-RNA at N7 position of guanine (m7G) to generate m7GpppA-RNA, (Fig. 2C, lanes 1 and 7). Either protein nsp10 or nsp16 was present but did not participate in the reaction. rep-rep-93 3725356 N/A 3725363 unspecified role unspecified role neutral component neutral component meres 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3725359 stimulator stimulator neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 3725360 unspecified role unspecified role 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 2D, 4B All four truncated Nsp10 proteins attenuated the SAM binding affinity of nsp16. rep-rep-96 3725356 N/A 3725359 competitor competitor competitor neutral component neutral component n-terminus 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false C-terminus amidated residue amidated residue certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3725363 unspecified role unspecified role neutral component neutral component meres 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3725359 stimulator stimulator neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 3725360 unspecified role unspecified role 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 4B rep-rep-94 3725357 N/A 3725359 ancillary ancillary neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 3725360 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 false ecoli Escherichia coli (strain K12) N/A 3725361 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 false 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 false in vitro In vitro methylation methylation reaction methylation false false false 4A rep-rep-95 3725357 N/A 3725359 competitor competitor competitor neutral component neutral component n-terminus 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 false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false C-terminus amidated residue amidated residue certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3725359 ancillary ancillary neutral component neutral component purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 3725360 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 false ecoli Escherichia coli (strain K12) N/A 3725361 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 false 2′-O-ribose methylated residue carbohydrate chemical modification carbohydrate chemical modification carbohydrate chemical modification certain certain sequence position certain certain certain sequence position certain false in vitro In vitro methylation methylation reaction methylation false false false 3725357 4A, rep-rep-90 3725358 N/A 3725360 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed N/A 3725359 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 1