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/ Reconstitution and functional characterization of SARS-CoV-2 proofreading complex. Reconstitution and functional characterization of SARS-CoV-2 proofreading complex. Protein expression and purification 2021 imex curation Ma Z., Pourfarjam Y., Kim IK. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus kimiw@ucmail.uc.edu Only protein-protein interactions imex curation in vitro In vitro enzymatic study enzymatic study tag fluorescence tag visualisation by fluorescence tag fluorescence Ma Z., Pourfarjam Y., Kim IK. Virus - Publications including interactions involving viral proteins I created new experiment rather than entering as both Polymerase AND nuclease assays;although 4C lane 9 is shown in E-1 as well - as no RTC present. (Although this interaction could be transferred to E-2 -Polymerase assay) RNA proofreading assay: RNA polymerase activity resulting in the extension of the primer/template RNA with a A:A mismatch, which occurs only in the presence of SARS-CoV-2 nsp14-nsp10. There is some concurrent nsp14-nsp10 exonuclease activity displayed shown by the presence of degraded FAM-RNA. kimiw@ucmail.uc.edu Accepted 2021-SEP-22 AT 13:28 BST AT 13:28 BST by ORCHARD Protein expression and purification 2021 Only protein-protein interactions imex curation imex curation Reconstitution and functional characterization of SARS-CoV-2 proofreading complex. Reconstitution and functional characterization of SARS-CoV-2 proofreading complex. Protein expression and purification 2021 imex curation Ma Z., Pourfarjam Y., Kim IK. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus kimiw@ucmail.uc.edu Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Ma Z., Pourfarjam Y., Kim IK. Virus - Publications including interactions involving viral proteins kimiw@ucmail.uc.edu Accepted 2021-SEP-22 AT 13:28 BST AT 13:28 BST by ORCHARD Protein expression and purification 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 Reconstitution and functional characterization of SARS-CoV-2 proofreading complex. Reconstitution and functional characterization of SARS-CoV-2 proofreading complex. Protein expression and purification 2021 imex curation Ma Z., Pourfarjam Y., Kim IK. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus kimiw@ucmail.uc.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 Ma Z., Pourfarjam Y., Kim IK. Virus - Publications including interactions involving viral proteins kimiw@ucmail.uc.edu Accepted 2021-SEP-22 AT 13:28 BST AT 13:28 BST by ORCHARD Protein expression and purification 2021 Only protein-protein interactions imex curation imex curation Reconstitution and functional characterization of SARS-CoV-2 proofreading complex. Reconstitution and functional characterization of SARS-CoV-2 proofreading complex. Protein expression and purification 2021 imex curation Ma Z., Pourfarjam Y., Kim IK. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus kimiw@ucmail.uc.edu Only protein-protein interactions imex curation in vitro In vitro ribonuclease assay ribonuclease assay tag fluorescence tag visualisation by fluorescence tag fluorescence Ma Z., Pourfarjam Y., Kim IK. Virus - Publications including interactions involving viral proteins kimiw@ucmail.uc.edu Accepted 2021-SEP-22 AT 13:28 BST AT 13:28 BST by ORCHARD Protein expression and purification 2021 Only protein-protein interactions imex curation imex curation 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 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 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 3' utr_40-mer-rna template rna ribonucleic acid RNA rna chemical synthesis Chemical synthesis (Chemical synthesis) CUAUCCCCAUGUGAUUUUAAUAGCUUCUUAGGAGAAUGAC residues 29831-29870 from 3' UTR 3' utr_21-mer-rna primer rna ribonucleic acid RNA rna chemical synthesis Chemical synthesis (Chemical synthesis) GUCAUUCUCCUAAGAAGCUAA 29870 - 29851 3' UTR plus 3-prime extra "A" 3' utr_20-mer-rna primer rna ribonucleic acid RNA rna chemical synthesis Chemical synthesis (Chemical synthesis) GUCAUUCUCCUAAGAAGCUA 29870 - 29851 3' UTR 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 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 3'-utr_40mer-dna template ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) CTATCCCCATGTGATTTTAATAGCTTCTTAGGAGAATGAC 29831 -29870 20mer_rna primer-3' utr_40_mer_rna template-1 5 9 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29831-29870 chemical synthesis Chemical synthesis (Chemical synthesis) 11 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29870 - 29851 5' prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 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) 8 enzyme enzyme neutral component neutral component purified purified n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 25 ecoli Escherichia coli (strain K12) 19 17 rna cleavage rna cleavage 1D I thought it better not to enter the complex here as roles cannot be shown for each component. The addition of nsp10 dramatically stimulates the nuclease activity of nsp14, forming a functional nuclease. Mg2+ (CHEBI:18420) Mn2+ (CHEBI:29035) EDTA (CHEBI:42191) Zn2+ (CHEBI:29105) Ca2+ (CHEBI:29108) 3' utr_20_mer_rna primer-3' utr_40_mer_rna template-1 4 11 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29870-29851 5' prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 31 enzyme enzyme neutral component neutral component purified purified C-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 33 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 35 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 37 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 39 ecoli Escherichia coli (strain K12) rna elongation rna strand elongation rna elongation 2B, 4B lane 2. Mg2+ (CHEBI:18420) Mn2+ (CHEBI:29035) EDTA (CHEBI:42191) Zn2+ (CHEBI:29105) Ca2+ (CHEBI:29108) 3' utr_20_mer_rna primer-3' utr_40_mer_rna template-2 5 9 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29831-29870 chemical synthesis Chemical synthesis (Chemical synthesis) 11 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29870-29851 5' prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 47 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 49 n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 25 ecoli Escherichia coli (strain K12) 44 42 rna cleavage rna cleavage 2A, 3A, 4A Mg2+ (CHEBI:18420) Mn2+ (CHEBI:29035) EDTA (CHEBI:42191) Zn2+ (CHEBI:29105) Ca2+ (CHEBI:29108) 3' utr_20_mer_rna primer-3' utr_40_mer_rna template-3 5 11 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29870-29851 5' prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 47 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 49 n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 25 ecoli Escherichia coli (strain K12) rna cleavage rna cleavage 3A 3' utr_20_mer_rna primer-3'_utr_40mer_dna template-1 5 14 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29831-29870 chemical synthesis Chemical synthesis (Chemical synthesis) 11 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29870-29851 5' prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 47 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 49 n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 25 ecoli Escherichia coli (strain K12) 60 62 rna cleavage rna cleavage 3A 3' utr_20_mer_rna primer-3' utr_40_mer_rna template-4 5 31 inhibitor inhibitor neutral component neutral component purified purified C-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 33 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 37 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 35 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 39 ecoli Escherichia coli (strain K12) 9 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29831-29870 chemical synthesis Chemical synthesis (Chemical synthesis) 11 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29870-29851 5' prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 47 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 49 n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 25 ecoli Escherichia coli (strain K12) 74 76 rna cleavage rna cleavage 4A Fig. 4A - I cannot say what the individual effects of Nsp 7/8/12 are, if any, and I dont think the authors do either. - so just entered the whole RTC. 3' utr_21_mer_rna primer-3' utr_40_mer_rna template-1 1 31 enzyme enzyme neutral component neutral component purified purified C-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 33 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 37 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 35 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 39 ecoli Escherichia coli (strain K12) 9 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29831-29870 chemical synthesis Chemical synthesis (Chemical synthesis) 10 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29870-29851 5' prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 47 enzyme regulator enzyme regulator 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 49 n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 25 ecoli Escherichia coli (strain K12) 90 88 rna elongation rna strand elongation rna elongation 4 B/C EDTA (CHEBI:64755) Mg2+ (CHEBI:18420) Ca2+ (CHEBI:29108) The addition of RTC partially protects the dsRNA from degradation by nsp14-nsp10. The RTC cannot extend this primer due to the A:A mismatch at the 3′ end of the primer. However, when SARS-CoV-2 nsp14-nsp10 is added to reactions, the full-length extension product (40-mer) starts to be detected. 3' utr_21_mer_rna primer-3' utr_40_mer_rna template-2 5 9 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29831-29870 chemical synthesis Chemical synthesis (Chemical synthesis) 10 enzyme target enzyme target substrate substrate neutral component neutral component 3-prime UTR region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 29870-29851 5' prime fluorescein fluorescein label fluorescein certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 47 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 49 n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 25 ecoli Escherichia coli (strain K12) 97 99 rna cleavage rna cleavage 4C Lane 9 rep-196 2 8 unspecified role unspecified role neutral component neutral component purified purified n-terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Supplementary figure 4 GST-nsp14 appears to be in equilibrium between different oligomeric states with the dimeric form as the predominant species. 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 6 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 7 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 110 107 107 109 110 108 109 108 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 nsp10-nsp14_sars2-1 SARS-CoV-2 3'-5' exoribonuclease proof-reading complex NSP10:NSP14 SARS-CoV-2 NSP14/NSP10 complex 8 enzyme enzyme nsp10 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role nsp14 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 112 111 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 The SARS-CoV-2 coronavirus 3'-5' exoribonuclease complex which strictly targets double-stranded RNA and can selectively remove a mismatched ribonucleotide at the 3'-end of a dsRNA substrate. Formation of the NSP10-NSP14 complex strongly enhances the exonuclease activity of the bifunctional NSP14 and enhances the fidelity of RNA synthesis by correcting nucleotide incorporation errors made by the RNA-dependent RNA polymerase. May bind to, and act with the nsp7/nsp8/nsp12 polymerase complex (CPX-5742). Proof-reading exonuclease activity may explain why coronaviruses have the largest RNA genomes known to date. Complex formation does not appear to effect the C-terminal N7-methyltransferase activity of NSP14 involved in RNA cap modification. 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. NSP10 forms a homododecamer in isolation. A flexible N-terminal domain of NSP14 forms the NSP10 docking site and induces conformational changes in the NSP14 that modulate the distance between the catalytic residues. New version remark: PMID:29279395 describes nsp14 as a 3'-5'-exoribonuclease. See also review PMID:31440227. Description, properties and xrefs updated and GO added as per Sandra's addition on SARS-CoV entry EBI-25496027/CPX-5707. Also updated this entry to "by orthology" as SARS-CoV has exp-evidence. Accepted 2020-APR-20 by ORCHARD coronavirus