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/ SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. Cellular & molecular immunology 2020 imex curation Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus taosc@sjtu.edu.cn 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins taosc@sjtu.edu.cn Accepted 2020-SEP-02 AT 08:50 BST AT 08:50 BST by MGT Cellular & molecular immunology 2020 Only protein-protein interactions imex curation imex curation SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. Cellular & molecular immunology 2020 imex curation Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus taosc@sjtu.edu.cn 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro bio-layer interferometry bio-layer interferometry bli bli predetermined predetermined participant predetermined Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins Authors have stated that TOM70 was not biotinlylated, it was just a purified protein. taosc@sjtu.edu.cn Accepted 2020-SEP-02 AT 08:50 BST AT 08:50 BST by MGT Cellular & molecular immunology 2020 Only protein-protein interactions imex curation imex curation Received response from authors, mentioning TOM70 was not biotinylated, i removed the tags and rest of the suggested executed. SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. Cellular & molecular immunology 2020 imex curation Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus taosc@sjtu.edu.cn 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins taosc@sjtu.edu.cn Accepted 2020-SEP-02 AT 08:50 BST AT 08:50 BST by MGT Cellular & molecular immunology 2020 Only protein-protein interactions imex curation imex curation all changes done..I had missed colocalization of TOM70 with SARS-CoV ORF9 previously, now added both..please have a look at them.. SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. Cellular & molecular immunology 2020 imex curation Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus taosc@sjtu.edu.cn 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down sequence tag sequence tag identification sequence tag Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins taosc@sjtu.edu.cn Accepted 2020-SEP-02 AT 08:50 BST AT 08:50 BST by MGT Cellular & molecular immunology 2020 Only protein-protein interactions imex curation imex curation host organism corrected.. SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. SARS-CoV-2 Orf9b suppresses type I interferon responses by targeting TOM70. Cellular & molecular immunology 2020 imex curation Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus taosc@sjtu.edu.cn 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight by staining molecular weight estimation by staining weight by staining Jiang HW., Zhang HN., Meng QF., Xie J., Li Y., Chen H., Zheng YX., Wang XN., Qi H., Zhang J., Wang PH., Han ZG., Tao SC. Virus - Publications including interactions involving viral proteins taosc@sjtu.edu.cn Accepted 2020-SEP-02 AT 08:50 BST AT 08:50 BST by MGT Cellular & molecular immunology 2020 Only protein-protein interactions imex curation imex curation done.. zfr_human Zinc finger RNA-binding protein ZFR M-phase phosphoprotein homolog protein protein human Homo sapiens Human MIPICPVVSFTYVPSRLGEDAKMATGNYFGFTHSGAAAAAAAAQYSQQPASGVAYSHPTTVASYTVHQAPVAAHTVTAAYAPAAATVAVARPAPVAVAAAATAAAYGGYPTAHTATDYGYTQRQQEAPPPPPPATTQNYQDSYSYVRSTAPAVAYDSKQYYQQPTATAAAVAAAAQPQPSVAETYYQTAPKAGYSQGATQYTQAQQTRQVTAIKPATPSPATTTFSIYPVSSTVQPVAAAATVVPSYTQSATYSTTAVTYSGTSYSGYEAAVYSAASSYYQQQQQQQKQAAAAAAAAAATAAWTGTTFTKKAPFQNKQLKPKQPPKPPQIHYCDVCKISCAGPQTYKEHLEGQKHKKKEAALKASQNTSSSNSSTRGTQNQLRCELCDVSCTGADAYAAHIRGAKHQKVVKLHTKLGKPIPSTEPNVVSQATSSTAVSASKPTASPSSIAANNCTVNTSSVATSSMKGLTTTGNSSLNSTSNTKVSAVPTNMAAKKTSTPKINFVGGNKLQSTGNKAEDIKGTECVKSTPVTSAVQIPEVKQDTVSEPVTPASLAALQSDVQPVGHDYVEEVRNDEGKVIRFHCKLCECSFNDPNAKEMHLKGRRHRLQYKKKVNPDLQVEVKPSIRARKIQEEKMRKQMQKEEYWRRREEEERWRMEMRRYEEDMYWRRMEEEQHHWDDRRRMPDGGYPHGPPGPLGLLGVRPGMPPQPQGPAPLRRPDSSDDRYVMTKHATIYPTEEELQAVQKIVSITERALKLVSDSLSEHEKNKNKEGDDKKEGGKDRALKGVLRVGVLAKGLLLRGDRNVNLVLLCSEKPSKTLLSRIAENLPKQLAVISPEKYDIKCAVSEAAIILNSCVEPKMQVTITLTSPIIREENMREGDVTSGMVKDPPDVLDRQKCLDALAALRHAKWFQARANGLQSCVIIIRILRDLCQRVPTWSDFPSWAMELLVEKAISSASSPQSPGDALRRVFECISSGIILKGSPGLLDPCEKDPFDTLATMTDQQREDITSSAQFALRLLAFRQIHKVLGMDPLPQMSQRFNIHNNRKRRRDSDGVDGFEAEGKKDKKDYDNF 3E43654E7134E9FC rabp2_human Cellular retinoic acid-binding protein 2 CRABP2 Cellular retinoic acid-binding protein II protein protein human Homo sapiens Human MPNFSGNWKIIRSENFEELLKVLGVNVMLRKIAVAAASKPAVEIKQEGDTFYIKTSTTVRTTEINFKVGEEFEEQTVDGRPCKSLVKWESENKMVCEQKLLKGEGPKTSWTRELTNDGELILTMTADDVVCTRVYVRE 982833700775B377 marcs_human Myristoylated alanine-rich C-kinase substrate Protein kinase C substrate, 80 kDa protein, light chain MARCKS MACS PRKCSL protein protein human Homo sapiens Human MGAQFSKTAAKGEAAAERPGEAAVASSPSKANGQENGHVKVNGDASPAAAESGAKEELQANGSAPAADKEEPAAAGSGAASPSAAEKGEPAAAAAPEAGASPVEKEAPAEGEAAEPGSPTAAEGEAASAASSTSSPKAEDGATPSPSNETPKKKKKRFSFKKSFKLSGFSFKKNKKEAGEGGEAEAPAAEGGKDEAAGGAAAAAAEAGAASGEQAAAPGEEAAAGEEGAAGGDPQEAKPQEAAVAPEKPPASDETKAAEEPSKVEEKKAEEAGASAAACEAPSAAGPGAPPEQEAAPAEEPAAAAASSACAAPSQEAQPECSPEAPPAEAAE 0AB247801EF8FCBF orf9b_sars2 ORF9b protein Accessory protein 9b ORF-9b Protein 9b 9b protein protein SARS-CoV-2 SARS-CoV-2 SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2 2019-nCoV MDPKISEMHPALRLVDPQIQLAVTRMENAVGRDQNNVGPKVYPIILRLGSPLSLNMARKTLNSLEDKAFQLTPIAVQMTKLATTEELPDEFVVVTVK orf9b_sars ORF9b protein Protein 9b 9b Accessory protein 9b ORF-9b protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus MDPNQTNVVPPALHLVDPQIQLTITRMEDAMGQGQNSADPKVYPIILRLGSQLSLSMARRNLDSLEARAFQSTPIVVQMTKLATTEELPDEFVVVTAK 5166F22872915FCB tom70_human Mitochondrial import receptor subunit TOM70 TOMM70 KIAA0719 TOM70 TOMM70A Translocase of outer membrane 70 kDa subunit Translocase of outer mitochondrial membrane protein 70 Mitochondrial precursor proteins import receptor protein protein human Homo sapiens Human MAASKPVEAAVVAAAVPSSGSGVGGGGTAGPGTGGLPRWQLALAVGAPLLLGAGAIYLWSRQQRRREARGRGDASGLKRNSERKTPEGRASPAPGSGHPEGPGAHLDMNSLDRAQAAKNKGNKYFKAGKYEQAIQCYTEAISLCPTEKNVDLSTFYQNRAAAFEQLQKWKEVAQDCTKAVELNPKYVKALFRRAKAHEKLDNKKECLEDVTAVCILEGFQNQQSMLLADKVLKLLGKEKAKEKYKNREPLMPSPQFIKSYFSSFTDDIISQPMLKGEKSDEDKDKEGEALEVKENSGYLKAKQYMEEENYDKIISECSKEIDAEGKYMAEALLLRATFYLLIGNANAAKPDLDKVISLKEANVKLRANALIKRGSMYMQQQQPLLSTQDFNMAADIDPQNADVYHHRGQLKILLDQVEEAVADFDECIRLRPESALAQAQKCFALYRQAYTGNNSSQIQAAMKGFEEVIKKFPRCAEGYALYAQALTDQQQFGKADEMYDKCIDLEPDNATTYVHKGLLQLQWKQDLDRGLELISKAIEIDNKCDFAYETMGTIEVQRGNMEKAIDMFNKAINLAKSEMEMAHLYSLCDAAHAQTEVAKKYGLKPPTL 5AAF5CAAA8582480 rap1b_human Ras-related protein Rap-1b OK/SW-cl.11 RAP1B GTP-binding protein smg p21B protein protein human Homo sapiens Human MREYKLVVLGSGGVGKSALTVQFVQGIFVEKYDPTIEDSYRKQVEVDAQQCMLEILDTAGTEQFTAMRDLYMKNGQGFALVYSITAQSTFNDLQDLREQILRVKDTDDVPMILVGNKCDLEDERVVGKEQGQNLARQWNNCAFLESSAKSKINVNEIFYDLVRQINRKTPVPGKARKKSSCQLL DIP protein P61224 original sequence version: 116 DIP protein P61224 original sequence version: 107 CE976895E5965224 orf9b_sars2-tomm70-1 1 9 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 1a orf9b_sars2-tomm70-2 2 9 unspecified role unspecified role bait bait purified purified c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 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 ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1b Avi-tag sequence allows the target to be biotinylated enzymatically BirA crabp2-1 4 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. 7 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. 11 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. 9 unspecified role unspecified role bait bait purified purified c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 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 ecoli Escherichia coli (strain K12) association association Supplementary Table 1, SF1a orf9b_sars2-tomm70-3 5 11 unspecified role unspecified role prey prey 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait purified purified c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 1e orf9b_sars2-tomm70-4 3 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization Figs. 1f, Supplementary Fig 2a orf9b_sars2-tomm70-5 3 9 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range colocalization colocalization Fig. 1g TOM70ΔTM overexpression partially disrupted the association of SARS-CoV2 Orf9b with mitochondria orf9b_sars2-tomm70-6 5 11 unspecified role unspecified role prey prey 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 c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait purified purified c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Supplementary Fig. 1b orf9b_cvhsa-tomm70-1 2 11 unspecified role unspecified role prey prey purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Kd was measured for increasing concentrations of TOM70 at 275, 550, 1100 nM ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role bait bait 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 c-terminus biotin tag biotin tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Supplementary Fig. 1c-d Avi-tag sequence allows the target to be biotinylated enzymatically BirA orf9b_cvhsa-orf9b_sars2-1 3 11 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell n-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Supplementary Fig 2b TOM70ΔTM overexpression partially disrupted the association of SARS-CoV Orf9b with mitochondria orf9b_cvhsa-tomm70-2 3 11 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell 10 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell c-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figs. 1f, Supplementary Fig 2a