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/ Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. J. Biol. Chem. (0021-9258) 2006 imex curation Xu Z., Sproul A., Wang W., Kukekov N., Greene LA. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology zhxu@genetics.ac.cn 14-DEC-2006: Contacted by JYOTI. Mouse brain. Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Xu Z., Sproul A., Wang W., Kukekov N., Greene LA. Apoptosis - Interactions involving proteins with a function related to apoptosis Radioactive proteins were visualized by SDS-PAGE/autoradiography zhxu@genetics.ac.cn Accepted 2006-JUN-22 by DAVET. 14-DEC-2006: Contacted by JYOTI. Mouse brain. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. J. Biol. Chem. (0021-9258) 2006 imex curation Xu Z., Sproul A., Wang W., Kukekov N., Greene LA. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology zhxu@genetics.ac.cn 14-DEC-2006: Contacted by JYOTI. Mouse brain. Only protein-protein interactions 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Xu Z., Sproul A., Wang W., Kukekov N., Greene LA. Apoptosis - Interactions involving proteins with a function related to apoptosis zhxu@genetics.ac.cn Accepted 2006-JUN-22 by DAVET. 14-DEC-2006: Contacted by JYOTI. Mouse brain. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. J. Biol. Chem. (0021-9258) 2006 imex curation Xu Z., Sproul A., Wang W., Kukekov N., Greene LA. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology zhxu@genetics.ac.cn 14-DEC-2006: Contacted by JYOTI. Mouse brain. Only protein-protein interactions rat-pc12 Rattus norvegicus PC12 cells pc12 Rat adrenal pheochromocytoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Xu Z., Sproul A., Wang W., Kukekov N., Greene LA. Apoptosis - Interactions involving proteins with a function related to apoptosis zhxu@genetics.ac.cn Accepted 2006-JUN-22 by DAVET. 14-DEC-2006: Contacted by JYOTI. Mouse brain. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. J. Biol. Chem. (0021-9258) 2006 imex curation Xu Z., Sproul A., Wang W., Kukekov N., Greene LA. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology zhxu@genetics.ac.cn 14-DEC-2006: Contacted by JYOTI. Mouse brain. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Xu Z., Sproul A., Wang W., Kukekov N., Greene LA. Apoptosis - Interactions involving proteins with a function related to apoptosis zhxu@genetics.ac.cn Accepted 2006-JUN-22 by DAVET. 14-DEC-2006: Contacted by JYOTI. Mouse brain. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation sia1a_mouse E3 ubiquitin-protein ligase SIAH1A Seven in absentia homolog 1a RING-type E3 ubiquitin transferase SIAH1A Siah1a protein protein mouse Mus musculus Mouse MSRQTATALPTGTSKCPPSQRVPALTGTTASNNDLASLFECPVCFDYVLPPILQCQSGHLVCSNCRPKLTCCPTCRGPLGSIRNLAMEKVANSVLFPCKYASSGCEITLPHTEKAEHEELCEFRPYSCPCPGASCKWQGSLDAVMPHLMHQHKSITTLQGEDIVFLATDINLPGAVDWVMMQSCFGFHFMLVLEKQEKYDGHQQFFAIVQLIGTRKQAENFAYRLELNGHRRRLTWEATPRSIHEGIATAIMNSDCLVFDTSIAQLFAENGNLGINVTISMC DIP protein P61092 original sequence version: 36 852EADC5DD4A4FFA siah1_human E3 ubiquitin-protein ligase SIAH1 Seven in absentia homolog 1 Siah-1a SIAH1 HUMSIAH RING-type E3 ubiquitin transferase SIAH1 protein protein human Homo sapiens Human MSRQTATALPTGTSKCPPSQRVPALTGTTASNNDLASLFECPVCFDYVLPPILQCQSGHLVCSNCRPKLTCCPTCRGPLGSIRNLAMEKVANSVLFPCKYASSGCEITLPHTEKADHEELCEFRPYSCPCPGASCKWQGSLDAVMPHLMHQHKSITTLQGEDIVFLATDINLPGAVDWVMMQSCFGFHFMLVLEKQEKYDGHQQFFAIVQLIGTRKQAENFAYRLELNGHRRRLTWEATPRSIHEGIATAIMNSDCLVFDTSIAQLFAENGNLGINVTISMC DIP protein Q8IUQ4 original sequence version: 132 FA0698D0DC1B0A15 sia1b_mouse E3 ubiquitin-protein ligase SIAH1B Siah1b Seven in absentia homolog 1b RING-type E3 ubiquitin transferase SIAH1B protein protein mouse Mus musculus Mouse MSRQAATALSTGTSKCPPSQRVPALTDTTASNNDLASLFECPVCFDYVLPPILQCQSGHLVCSNCRPKLTCCPTCRGPLGSIRNLAVEKVANSVLFPCKYSASGCEITLPHTKKAEHEELCEFRPYSCPCPGASCKWQGSLDAVMPHLMHQHKSITTLQGEDIVFLATDINLPGAVDWVMMQSCFGFHFMLVLEKQEKYDGHQQFFAIVQLIGTRKQAENFAYRLELNGHRRRLTWEATPRSIHEGIATAIMNSDCLVFDTSIAQLFAENGNLGINVTISMC A09F5D3DEEB39AC2 sh3r1_rat E3 ubiquitin-protein ligase SH3RF1 Posh1 Sh3rf1 Posh Sh3md2 SH3 domain-containing RING finger protein 1 Plenty of SH3s SH3 multiple domains protein 2 RING-type E3 ubiquitin transferase SH3RF1 protein protein rat Rattus norvegicus (Rat) Rat MDESALLDLLECPVCLERLDASAKVLPCQHTFCKRCLLGIVGSRNELRCPECRTLVGSGVDELPSNILLVRLLDGIKQRPWKPGPGGGGSTTCTNVLRAQGSTVVNCGSKDLQSPQCGQQPRVQAWSPPVRGIPQLPCAKALYNYEGKEPGDLKFSKGDIIILRRQVDENWYHGEVNGVHGFFPTNFVQIIKPLPQPPPQCKALYDFEVKDKEADKDCLPFAKDDVLTVIRRVDENWAEGMLADKIGIFPISYVEFNSAAKQLIEWDKPPVPGVDTAECPSATAAQSSSASKHSDTKKNTRKRHSFTSLTMANKSSQASQNRHSMEISPPVLISSSNPTAAARISELSGLSCSAPSQVHISTTGLIVTPPPSSPVTTGPSFTFPTDVPYQAALGTMNPPLPPPPLLATTVLASTPSGATAAAVAAAAAAVAAGVGPRPAVGSTEQIAHLRPQTRPSVYVAIYPYTPRKEDELELRKGEMFLVFERCQDGWYKGTSMHTSKIGVFPGNYVAPVTRAVTNASQAKVPMSTAGQASRGVTMVSPSTAGGPAQKPQGNGVAGNPSVVPTAVVSAAHIQTSPQAKVLLHMTGQMTVNQARNAVRTVAAHNQERPTAAVTPIQVQNAACIGPASVGLPHHSLASQPLPPMVGPAAHIAAVNINRTSVPLACAAGASSLASPNMTTAALETEPSGRTVTILPGLPTSPESAASACGNSSAVKPDKDSKKEKKGLLKLLSGASTKRKPRVSPPASPTLDVELGSGEVPLQGAVGPELPLGGVHGRVGSCPTDGDGPVAAGTAALAQDAFHRKTSSLDSAVPIAPPPRQACSSLGPVMNEARPVVCERHRVVVSYPPQSEAELELKEGDIVFVHKKREDGWFKGTLQRNGKTGLFPGSFVENI E7CB1B7E6330C62D siah2_mouse E3 ubiquitin-protein ligase SIAH2 Siah2 Seven in absentia homolog 2 RING-type E3 ubiquitin transferase SIAH2 protein protein mouse Mus musculus Mouse MSRPSSTGPSANKPCSKQPPPPQTPHAPSPAAPPAAATISAAGPGSSAVPAAAAVISGPGAGGGADPVSPQHHELTSLFECPVCFDYVLPPILQCQAGHLVCNQCRQKLSCCPTCRGALTPSIRNLAMEKVASAVLFPCKYATTGCSLTLHHTEKPEHEDICEYRPYSCPCPGASCKWQGSLEAVMSHLMHAHKSITTLQGEDIVFLATDINLPGAVDWVMMQSCFGHHFMLVLEKQEKYEGHQQFFAIVLLIGTRKQAENFAYRLELNGNRRRLTWEATPRSIHDGVAAAIMNSDCLVFDTAIAHLFADNGNLGINVTISTCCQ 1CB73AD3B4C9982F sh3r1_mouse E3 ubiquitin-protein ligase SH3RF1 Posh1 Sh3rf1 Kiaa1494 Posh SH3 domain-containing RING finger protein 1 Plenty of SH3s SH3 multiple domains protein 2 Sh3md2 RING-type E3 ubiquitin transferase SH3RF1 protein protein mouse Mus musculus Mouse MDESALLDLLECPVCLERLDASAKVLPCQHTFCKRCLLGIVGSRNELRCPECRTLVGSGVDELPSNILLVRLLDGIKQRPWKPGPGGGGGTTCTNTLRAQGSTVVNCGSKDLQSSQCGQQPRVQAWSPPVRGIPQLPCAKALYNYEGKEPGDLKFSKGDIIILRRQVDENWYHGEVSGVHGFFPTNFVQIIKPLPQPPPQCKALYDFEVKDKEADKDCLPFAKDDVLTVIRRVDENWAEGMLADKIGIFPISYVEFNSAAKQLIEWDKPPVPGVDTAECPSATAQSTSASKHPDTKKNTRKRHSFTSLTMANKSSQGSQNRHSMEISPPVLISSSNPTAAARISELSGLSCSAPSQVHISTTGLIVTPPPSSPVTTGPAFTFPSDVPYQAALGSMNPPLPPPPLLAATVLASTPSGATAAVAAAAAAAAAAGMGPRPVMGSSEQIAHLRPQTRPSVYVAIYPYTPRKEDELELRKGEMFLVFERCQDGWYKGTSMHTSKIGVFPGNYVAPVTRAVTNASQAKVSMSTAGQASRGVTMVSPSTAGGPTQKPQGNGVAGNPSVVPTAVVSAAHIQTSPQAKVLLHMSGQMTVNQARNAVRTVAAHSQERPTAAVTPIQVQNAACLGPASVGLPHHSLASQPLPPMAGPAAHGAAVSISRTNAPMACAAGASLASPNMTSAMLETEPSGRTVTILPGLPTSPESAASACGNSSAGKPDKDSKKEKKGLLKLLSGASTKRKPRVSPPASPTLDVELGAGEAPLQGAVGPELPLGGSHGRVGSCPTDGDGPVAAGTAALAQDAFHRKTSSLDSAVPIAPPPRQACSSLGPVMNEARPVVCERHRVVVSYPPQSEAELELKEGDIVFVHKKREDGWFKGTLQRNGKTGLFPGSFVENI D92071116F4D6EBE siah1_rat E3 ubiquitin-protein ligase SIAH1 Siah1 Siah1a Seven in absentia homolog 1 Siah-1a RING-type E3 ubiquitin transferase SIAH1 protein protein rat Rattus norvegicus (Rat) Rat MSRQTATALPTGTSKCPPSQRVPALTGTTASNNDLASLFECPVCFDYVLPPILQCQSGHLVCSNCRPKLTCCPTCRGPLGSIRNLAMEKVANSVLFPCKYASSGCEITLPHTEKAEHEELCEFRPYSCPCPGASCKWQGSLDAVMPHLMHQHKSITTLQGEDIVFLATDINLPGAVDWVMMQSCFGFHFMLVLEKQEKYDGHQQFFAIVQLIGTRKQAENFAYRLELNGHRRRLTWEATPRSIHEGIATAIMNSDCLVFDTSIAQLFAENGNLGINVTISMC DIP protein Q920M9 original sequence version: 104 852EADC5DD4A4FFA sh3md2-siah1a-1 2 10 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association Full-length POSH was cloned into a bait vector, and a total of ~2 x 106 independent transformants from a mouse brain library were analyzed. Fifty-two strong to very strong positive clones were observed, and their identities were established by sequencing. Fourteen clones corresponded to Siah1a, two to Siah1b, and three to Siah2 sh3md2-siah1b-2 2 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Full-length POSH was cloned into a bait vector, and a total of ~2 x 106 independent transformants from a mouse brain library were analyzed. Fifty-two strong to very strong positive clones were observed, and their identities were established by sequencing. Fourteen clones corresponded to Siah1a, two to Siah1b, and three to Siah2 sh3md2-siah2-1 2 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Full-length POSH was cloned into a bait vector, and a total of ~2 x 106 independent transformants from a mouse brain library were analyzed. Fifty-two strong to very strong positive clones were observed, and their identities were established by sequencing. Fourteen clones corresponded to Siah1a, two to Siah1b, and three to Siah2 sh3md2-siah1-1 4 6 unspecified role unspecified role bait bait Q8IUQ4:p.[Cys41Ser;Cys44Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Wed May 02 18:10:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys41ser,cys44ser' Tue Jun 30 16:28:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys41Ser;Cys44Ser]' region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range certain certain sequence position certain certain certain sequence position certain range range physical association physical association used co-immunoprecipitation to confirm that SIAH1 and POSH interact in mammalian cells 1A RING finger domain of Sh3md2 deleted and shown not to be essential for interaction. Domain boundaries of deletion mutant not given, therefore estimated using ProSite domain boundaries. sh3md2-siah1-2 4 10 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range certain certain sequence position certain certain certain sequence position certain range range 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q8IUQ4:p.[Cys41Ser;Cys44Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain C S certain certain sequence position certain certain certain sequence position certain C S Wed May 02 20:04:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys41ser,cys44ser' Tue Jun 30 16:58:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Cys41Ser;Cys44Ser]' physical association physical association used co-immunoprecipitation to confirm that SIAH1 and POSH interact in mammalian cells 1B RING finger domain of Sh3md2 deleted and shown not to be essential for interaction. Domain boundaries of deletion mutant not given, therefore estimated using ProSite domain boundaries. sh3md2-siah1-3 4 6 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 10 unspecified role unspecified role prey prey region delta ring sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range certain certain sequence position certain certain certain sequence position certain range range region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The C-terminal 52 amino acids of SIAH1 are required for dimerization and auto-destabilization (5, 29). We found that, despite its greatly enhanced stability and consequent elevated expression, SIAH1 with these residues deleted (SIAH1 {Delta}C2) was severely compromised for interaction with POSH 1C Exact amino acid residues removed from deltaRING construct of POSH (O70254) not given in entry, so taken from ProSite pattern PS00518. sh3md2-siah1-4 1 6 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 10 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction the ability of GST-POSH fusion proteins to pull down SIAH1 1D sh3md2-siah1-5 3 11 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association endogenous SIAH1 and POSH interact with each other. PC12 cells 1E Camptothecin - a pro-apoptotic signal sh3md2-siah1-6 3 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association endogenous SIAH1 and POSH interact with each other. PC12 cells 1E Camptothecin - a pro-apoptotic signal sh3md2-siah1-7 4 6 unspecified role unspecified role prey prey c-terminal region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait region delta ring sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range certain certain sequence position certain certain certain sequence position certain range range region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The C-terminal 52 amino acids of SIAH1 are required for dimerization and auto-destabilization (5, 29). We found that, despite its greatly enhanced stability and consequent elevated expression, SIAH1 with these residues deleted (SIAH1 {Delta}C2) was severely compromised for interaction with POSH 1C Exact amino acid residues removed from deltaRING construct of POSH (O70254) not given in entry, so taken from ProSite pattern PS00518.