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/ Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. J. Biol. Chem. (0021-9258) 2008 Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA human-rko Homo sapiens colorectal carcinoma cell line rko Human colorectal carcinoma cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr Accepted 2008-NOV-6 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA J. Biol. Chem. (0021-9258) 2008 Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. J. Biol. Chem. (0021-9258) 2008 Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA 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 anti tag western anti tag western blot anti tag western Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr Accepted 2008-NOV-6 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA J. Biol. Chem. (0021-9258) 2008 orchard Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. J. Biol. Chem. (0021-9258) 2008 Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr Accepted 2008-NOV-6 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA J. Biol. Chem. (0021-9258) 2008 Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. J. Biol. Chem. (0021-9258) 2008 Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA 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 Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr Accepted 2008-NOV-6 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA J. Biol. Chem. (0021-9258) 2008 Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. Ubiquitination and degradation of homeodomain-interacting protein kinase 2 by WD40 repeat/SOCS box protein WSB-1. J. Biol. Chem. (0021-9258) 2008 Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr 14-Nov-2008: Contacted by JYOTI. Mouse embryonic day 11 cDNA 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 Choi DW., Seo YM., Kim EA., Sung KS., Ahn JW., Park SJ., Lee SR., Choi CY. Apoptosis - Interactions involving proteins with a function related to apoptosis The source of HIPK2 was taken from PMID 9748262. choicy@skku.ac.kr Accepted 2008-NOV-6 by ORCHARD. Mouse embryonic day 11 cDNA J. Biol. Chem. (0021-9258) 2008 hipk2_mouse Homeodomain-interacting protein kinase 2 Hipk2 Nbak1 Stank Nuclear body-associated kinase 1 Sialophorin tail-associated nuclear serine/threonine-protein kinase protein protein mouse Mus musculus Mouse MAPVYEGMASHVQVFSPHTLQSSAFCSVKKLKVEPSSNWDMTGYGSHSKVYSQSKNIPPSQPASTTVSTSLPIPNPSLPYEQTIIFPGSTGHIVVTSASSTSVTGQVLGGPHNLMRRSTVSLLDTYQKCGLKRKSEEIENTSSVQIIEEHPPMIQNNASGATVATATTSTATSKNSGSNSEGDYQLVQHEVLCSMTNTYEVLEFLGRGTFGQVVKCWKRGTNEIVAIKILKNHPSYARQGQIEVSILARLSTESADDYNFVRAYECFQHKNHTCLVFEMLEQNLYDFLKQNKFSPLPLKYIRPVLQQVATALMKLKSLGLIHADLKPENIMLVDPSRQPYRVKVIDFGSASHVSKAVCSTYLQSRYYRAPEIILGLPFCEAIDMWSLGCVIAELFLGWPLYPGASEYDQIRYISQTQGLPAEYLLSAGTKTTRFFNRDTDSPYPLWRLKTPDDHEAETGIKSKEARKYIFNCLDDMAQVNMTTDLEGSDMLVEKADRREFIDLLKKMLTIDADKRVTPIETLNHPFVTMTHLLDFPHSAHVKSCFQNMEICKRRVNMYDTVNQSKTPFITHVAPSTSTNLTMTFNNQLTTVHNQAPTTSSATLSLANPEVSILNYQSALYQPSAASMAAVAPRSMPLQTGTAQICARPDPFQQALIVCPPGFQGLQASPSKHAGYSVRMENAVPIVTQAPGAQPLQIQPGLLAQQAWPGGAQQILLPPAWQQLTGVATHTSVQHAAVIPETMAGTQQLADWRNTHAHGSHYNPIMQQPALLTGHVTLPAAQPLNVGVAHVMRQQPTSTTSSRKSKQHQSSVRNVSTCEVTSSQAISSPQRSKRVKENTPPRCAMVHSSPACSTSVTCGWGDVASSTTRERQRQTIVIPDTPSPTVSVITISSDTDEEEEQKHAPTSTVSKQRKNVISCVTVHDSPYSDSSSNTSPYSVQQRTGHNGTNTLDTKGGLENHCTGNPRTIIVPPLKTQASEVLVECDSLGPAISASHHSSSFKSKSSSTVTSTSGHSSGSSSGAIAYRQQRPGPHFQQQQPLNLSQAQQHMAADRTGSHRRQQAYITPTMAQAPYTFPHNSPSHGTVHPHLAAAAHLPTQPHLYTYTAPTALGSTGTVAHLVASQGSARHTVQHTAYPASIVHQVPVSMGPRVLPSPTIHPSQYPAQFAHQTYISASPASTVYTGYPLSPAKVNQYPYI 5C863BE377F3AAEF 5C863BE377F3AAEF wsb1_mouse WD repeat and SOCS box-containing protein 1 Wsb1 protein protein mouse Mus musculus Mouse MASFPPRVNEKEIVRSRTIGELLAPAAPFDKKCGGENWTVAFAPDGSYFAWSQGYRIVKLVPWSQCRKNFLLHGSKNVTNSSCLKLARQNSNGGQKNKPPEHVIDCGDIVWSLAFGSSVPEKQSRCVNIEWHRFRFGQDQLLLATGLNNGRIKIWDVYTGKLLLNLVDHIEMVRDLTFAPDGSLLLVSASRDKTLRVWDLKDDGNMVKVLRAHQNWVYSCAFSPDCSMLCSVGASKAVFLWNMDKYTMIRKLEGHHHDVVACDFSPDGALLATASYDTRVYVWDPHNGDLLMEFGHLFPSPTPIFAGGANDRWVRAVSFSHDGLHVASLADDKMVRFWRIDEDCPVQVAPLSNGLCCAFSTDGSVLAAGTHDGSVYFWATPRQVPSLQHICRMSIRRVMSTQEVQKLPVPSKILAFLSYRG 4003D1FFE7A9A2EF hipk2_human Homeodomain-interacting protein kinase 2 HIPK2 protein protein human Homo sapiens Human MAPVYEGMASHVQVFSPHTLQSSAFCSVKKLKIEPSSNWDMTGYGSHSKVYSQSKNIPLSQPATTTVSTSLPVPNPSLPYEQTIVFPGSTGHIVVTSASSTSVTGQVLGGPHNLMRRSTVSLLDTYQKCGLKRKSEEIENTSSVQIIEEHPPMIQNNASGATVATATTSTATSKNSGSNSEGDYQLVQHEVLCSMTNTYEVLEFLGRGTFGQVVKCWKRGTNEIVAIKILKNHPSYARQGQIEVSILARLSTESADDYNFVRAYECFQHKNHTCLVFEMLEQNLYDFLKQNKFSPLPLKYIRPVLQQVATALMKLKSLGLIHADLKPENIMLVDPSRQPYRVKVIDFGSASHVSKAVCSTYLQSRYYRAPEIILGLPFCEAIDMWSLGCVIAELFLGWPLYPGASEYDQIRYISQTQGLPAEYLLSAGTKTTRFFNRDTDSPYPLWRLKTPDDHEAETGIKSKEARKYIFNCLDDMAQVNMTTDLEGSDMLVEKADRREFIDLLKKMLTIDADKRITPIETLNHPFVTMTHLLDFPHSTHVKSCFQNMEICKRRVNMYDTVNQSKTPFITHVAPSTSTNLTMTFNNQLTTVHNQAPSSTSATISLANPEVSILNYPSTLYQPSAASMAAVAQRSMPLQTGTAQICARPDPFQQALIVCPPGFQGLQASPSKHAGYSVRMENAVPIVTQAPGAQPLQIQPGLLAQQAWPSGTQQILLPPAWQQLTGVATHTSVQHATVIPETMAGTQQLADWRNTHAHGSHYNPIMQQPALLTGHVTLPAAQPLNVGVAHVMRQQPTSTTSSRKSKQHQSSVRNVSTCEVSSSQAISSPQRSKRVKENTPPRCAMVHSSPACSTSVTCGWGDVASSTTRERQRQTIVIPDTPSPTVSVITISSDTDEEEEQKHAPTSTVSKQRKNVISCVTVHDSPYSDSSSNTSPYSVQQRAGHNNANAFDTKGSLENHCTGNPRTIIVPPLKTQASEVLVECDSLVPVNTSHHSSSYKSKSSSNVTSTSGHSSGSSSGAITYRQQRPGPHFQQQQPLNLSQAQQHITTDRTGSHRRQQAYITPTMAQAPYSFPHNSPSHGTVHPHLAAAAAAAHLPTQPHLYTYTAPAALGSTGTVAHLVASQGSARHTVQHTAYPASIVHQVPVSMGPRVLPSPTIHPSQYPAQFAHQTYISASPASTVYTGYPLSPAKVNQYPYI DIP protein Q9H2X6 original sequence version: 132 6022D5710E8D2D93 wsb1_human WD repeat and SOCS box-containing protein 1 SOCS box-containing WD protein SWiP-1 WSB1 SWIP1 protein protein human Homo sapiens Human MASFPPRVNEKEIVRLRTIGELLAPAAPFDKKCGRENWTVAFAPDGSYFAWSQGHRTVKLVPWSQCLQNFLLHGTKNVTNSSSLRLPRQNSDGGQKNKPREHIIDCGDIVWSLAFGSSVPEKQSRCVNIEWHRFRFGQDQLLLATGLNNGRIKIWDVYTGKLLLNLVDHTEVVRDLTFAPDGSLILVSASRDKTLRVWDLKDDGNMMKVLRGHQNWVYSCAFSPDSSMLCSVGASKAVFLWNMDKYTMIRKLEGHHHDVVACDFSPDGALLATASYDTRVYIWDPHNGDILMEFGHLFPPPTPIFAGGANDRWVRSVSFSHDGLHVASLADDKMVRFWRIDEDYPVQVAPLSNGLCCAFSTDGSVLAAGTHDGSVYFWATPRQVPSLQHLCRMSIRRVMPTQEVQELPIPSKLLEFLSYRI 650D4942E97D4BED hipk2-wsb1 5 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey physical association physical association theCterminus of HIPK2-(503-1189) was utilized as bait for the screening of mouse embryonic match-maker cDNA libraries. Among the several cellular proteins that were identified and sequenced, a clone showed identity to WSB-1, a subunit of E3 ubiquitin ligase Author states that the C-terminus of HIPK2 terminates at 1189 - this does not agree with the UniProt protein sequence hipk2-wsb1-1 2 6 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 9 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The specific interactions between WSB-1 and HIPK2 were verified in the yeast two-hybrid and co-immunoprecipitation assays in cultured cells 2 C Q9Y6I7; Q9H2X6 transfection hipk2-wsb1-2 4 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range 7 unspecified role unspecified role prey prey physical association physical association The authors state that the C-terminus of HIPK2 (aa 503-1189) was used as bait. However, the UniProtKB entry for HIPK2 is 1196 aa in length. Thus approximate coordinates have been used for the range of HIPK2 in this interaction. 2 B The specific interactions between WSB-1 and HIPK2 were verified in the yeast two-hybrid hipk2-wsb1-3 3 6 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 n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Q9QZR5:p.Lys228Arg mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K R Wed May 02 19:47:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys228arg' Tue Jun 30 19:49:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys228Arg' in vitro In vitro 9 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 26 29 physical association physical association 2 D, 7 B, C GST pull-down analysis showed that WSB-1 interacted physically with HIPK2 Dephosphorylation of wild-type HIPK2 with alkaline phosphatase increased binding of HIPK2 to WSB-1 but not of the catalytically inactive HIPK2 K221R mutant. hipk2-wsb1-4 1 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 2 E The association of endogenous HIPK2 with WSB-1 was also verified via co-immunoprecipitation with anti-HIPK2 antibody followed by Western blotting using anti-WSB-1 antibody hipk2-wsb1-5 3 9 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 n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain undefined mutation mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:16:40 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'phe359asp362leu366' in vitro In vitro 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) physical association physical association 5 B GSTpulldown analysis with GST-HIPK2-(503-860) and Myc-tagged WSB-1 mutants showed that all of the deletion mutants of WSB-1 containing theWD40repeat interacted with HIPK2 hipk2-wsb1-6 2 6 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 9 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 5 C both N-terminal WSB-1 (aa 1-279, 1-5th WD40 repeats) and C-terminal WSB-1 (aa 200-421, 4-7th WD40 repeats and SOCS box) were associated with full-length HIPK2 in cultured cells Q9Y6I7; Q9QZR5 transfection hipk2-wsb1-7 2 6 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 9 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 physical association physical association 7 D The bindings of WSB-1 to both wild-type HIPK2 and catalytically inactive HIPK2 K221R mutant were obviously diminished following Adriamycin administration. Binding of WSB-1 to wild-type HIPK2 and kinase dead HIPK2 K221R was decreased after Adriamycin treatment. The effect was more pronounced with wild-type HIPK2. Q9Y6I7; Q9QZR5 transfection