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/ The enzymatic activity of the nsp14 exoribonuclease is critical for replication of MERS-CoV and SARS-CoV-2 The enzymatic activity of the nsp14 exoribonuclease is critical for replication of MERS-CoV and SARS-CoV-2 Journal of Virology 2020 imex curation Ogando N.S. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus E.J.Snijder@lumc.nl 05-Jan-2021: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro ribonuclease assay ribonuclease assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Ogando N.S. Virus - Publications including interactions involving viral proteins E.J.Snijder@lumc.nl Accepted 2020-DEC-07 AT 12:06 GMT AT 12:06 GMT by PPORRAS Journal of Virology 2020 Only protein-protein interactions imex curation imex curation The enzymatic activity of the nsp14 exoribonuclease is critical for replication of MERS-CoV and SARS-CoV-2 The enzymatic activity of the nsp14 exoribonuclease is critical for replication of MERS-CoV and SARS-CoV-2 Journal of Virology 2020 imex curation Ogando N.S. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus E.J.Snijder@lumc.nl 05-Jan-2021: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro radiolabeled methyl methyltransferase radiometric assay radiolabeled methyl predetermined predetermined participant predetermined Ogando N.S. Virus - Publications including interactions involving viral proteins E.J.Snijder@lumc.nl Accepted 2020-DEC-07 AT 12:06 GMT AT 12:06 GMT by PPORRAS Journal of Virology 2020 Only protein-protein interactions imex curation imex curation p0dtd1-pro_0000449628 Non-structural protein 10 rep ORF1ab polyprotein 1a-1b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV AGNATEVPANSTVLSFCAFAVDAAKAYKDYLASGGQPITNCVKMLCTHTGTGQAITVTPEANMDQESFGGASCCLYCRCHIDHPNPKGFCDLKGKYVQIPTTCANDPVGFTLKNTVCTVCGMWKGYGCSCDQLREPMLQ 4254 4392 5'-end (5'-triphospho-guanosine)-adenosine(3-) residue 5'-end (5'-triphospho-guanosine)-adenosine(3-) residue a 5'-end (5'-triphospho-guanosine)-adenosine residue small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) k9n7c7-pro_0000422451 Guanine-N7 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 SQIVTGLFKDCSRETSGLSPAYAPTYVSVDDKYKTSDELCVNLNLPANVPYSRVISRMGFKLDATVPGYPKLFITREEAVRQVRSWIGFDVEGAHASRNACGTNVPLQLGFSTGVNFVVQPVGVVDTEWGNMLTGIAARPPPGEQFKHLVPLMHKGAAWPIVRRRIVQMLSDTLDKLSDYCTFVCWAHGFELTSASYFCKIGKEQKCCMCNRRAAAYSSPLQSYACWTHSCGYDYVYNPFFVDVQQWGYVGNLATNHDRYCSVHQGAHVASNDAIMTRCLAIHSCFIERVDWDIEYPYISHEKKLNSCCRIVERNVVRAALLAGSFDKVYDIGNPKGIPIVDDPVVDWHYFDAQPLTRKVQQLFYTEDMASRFADGLCLFWNCNVPKYPNNAIVCRFDTRVHSEFNLPGCDGGSLYVNKHAFHTPAYDVSAFRDLKPLPFFYYSTTPCEVHGNGSMIEDIDYVPLKSAVCITACNLGGAVCRKHATEYREYMEAYNLVSASGFRLWCYKTFDIYNLWSTFTKVQ 5909 6432 D1A1D3C8610137A1 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 p0dtd1-pro_0000449631 Proofreading exoribonuclease nsp14 rep ORF1ab polyprotein 1a-1b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV AENVTGLFKDCSKVITGLHPTQAPTHLSVDTKFKTEGLCVDIPGIPKDMTYRRLISMMGFKMNYQVNGYPNMFITREEAIRHVRAWIGFDVEGCHATREAVGTNLPLQLGFSTGVNLVAVPTGYVDTPNNTDFSRVSAKPPPGDQFKHLIPLMYKGLPWNVVRIKIVQMLSDTLKNLSDRVVFVLWAHGFELTSMKYFVKIGPERTCCLCDRRATCFSTASDTYACWHHSIGFDYVYNPFMIDVQQWGFTGNLQSNHDLYCQVHGNAHVASCDAIMTRCLAVHECFVKRVDWTIEYPIIGDELKINAACRKVQHMVVKAALLADKFPVLHDIGNPKAIKCVPQADVEWKFYDAQPCSDKAYKIEELFYSYATHSDKFTDGVCLFWNCNVDRYPANSIVCRFDTRVLSNLNLPGCDGGSLYVNKHAFHTPAFDKSAFVNLKQLPFFYYSDSPCESHGKQVVSDIDYVPLKSATCITRCNLGGAVCRHHANEYRLYLDAYNMMISAGFSLWVYKQFDTYNLWNTFTRLQ 5926 6452 h4_rna rna ribonucleic acid RNA rna synthetic construct synthetic construct UGACGGCCCGGAAAACCGGGCC h4_rna-rep-1 1 5 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) 8 enzyme target enzyme target substrate substrate neutral component neutral component 5' 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain synthetic construct synthetic construct rna cleavage rna cleavage 7,8,9,10 Trace activity of Nsp14 alone was seen. h4_rna-rep-2 1 8 enzyme target enzyme target substrate substrate neutral component neutral component 5' 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain synthetic construct synthetic construct 6 stimulator stimulator 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 enzyme enzyme neutral component neutral component purified purified K9N7C7-PRO_0000422451:p.Asp90Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain D A K9N7C7-PRO_0000422451:p.Glu92Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain E A K9N7C7-PRO_0000422451:p.His268Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain H A K9N7C7-PRO_0000422451:p.Glu191Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain E A K9N7C7-PRO_0000422451:p.Asp273Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A K9N7C7-PRO_0000422451:p.Asp331Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain D A K9N7C7-PRO_0000422451:p.Arg310Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R A 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_0000422451:p.[Asp90Ala;Glu92Ala] mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A certain certain sequence position certain certain certain sequence position certain E A Thu Oct 29 22:18:47 GMT 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'K9N7C7-PRO_0000422451:p.AspD90Ala;Glu92Ala' K9N7C7-PRO_0000422451:p.Glu191Asp mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain E D K9N7C7-PRO_0000422451:p.His229Cys mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain H C ecoli Escherichia coli (strain K12) rna cleavage rna cleavage 7,8,9,10 MERS-CoV nsp14 ExoN activity was found to be stimulated by 425 nsp10 in a dose-dependent manner. MERS-CoV nsp14 ExoN activity was found to be stimulated by 425 nsp10 in a dose-dependent manner h4_rna-rep-3 1 8 enzyme target enzyme target substrate substrate neutral component neutral component 5' 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain synthetic construct synthetic construct 3 stimulator stimulator 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 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) rna cleavage rna cleavage 9 h4_rna-rep-4 1 8 enzyme target enzyme target substrate substrate neutral component neutral component 5' 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain synthetic construct synthetic construct 6 stimulator stimulator 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 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) rna cleavage rna cleavage 9 SARS Nsp14 +MERS Nsp10 h4_rna-rep-5 1 8 enzyme target enzyme target substrate substrate neutral component neutral component 5' 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain synthetic construct synthetic construct 3 stimulator stimulator 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 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 P0DTD1-PRO_0000449631:p.[Asp90Ala;Glu92Ala] mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain D A certain certain sequence position certain certain certain sequence position certain E A Thu Oct 29 22:17:56 GMT 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'K9N7C7-PRO_0000422451:p.AspD90Ala;Glu92Ala' ecoli Escherichia coli (strain K12) rna cleavage rna cleavage 9 MERS Nsp14 + SARS Nsp10 h4_rna-rep-6 1 7 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) 8 enzyme target enzyme target substrate substrate neutral component neutral component 5' 32p radiolabel 32p radiolabel P32 32P 32P P32 certain certain sequence position certain certain certain sequence position certain synthetic construct synthetic construct rna cleavage rna cleavage 9 Trace activity of Nsp14 alone was seen. 5'_end (5'_triphospho_guanosine)_adenosine(3_) residue-rep-2 2 4 enzyme target enzyme target substrate substrate neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 5 enzyme enzyme neutral component neutral component purified purified K9N7C7-PRO_0000422451:p.Asp90Ala 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 K9N7C7-PRO_0000422451:p.Glu92Ala 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 E A K9N7C7-PRO_0000422451:p.His268Ala 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 H A K9N7C7-PRO_0000422451:p.Glu191Ala 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 E A K9N7C7-PRO_0000422451:p.Asp273Ala 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 K9N7C7-PRO_0000422451:p.Asp331Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain D A K9N7C7-PRO_0000422451:p.Arg310Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain R A 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_0000422451:p.[Asp90Ala;Glu92Ala] 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 certain certain sequence position certain certain certain sequence position certain E A Thu Oct 29 22:43:10 GMT 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'K9N7C7-PRO_0000422451:p.AspD90Ala;Glu92Ala' K9N7C7-PRO_0000422451:p.Glu191Asp 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 E D K9N7C7-PRO_0000422451:p.His229Cys 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 H C ecoli Escherichia coli (strain K12) methylation methylation reaction methylation 11