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/ Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. J. Biol. Chem. (0021-9258) 2001 imex curation Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer lieberma@unix.temple.edu 21-Dec-2006: Contacted by JYOTI. Only protein-protein interactions 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 western blot western blot Immuno blot Immuno blot Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis lieberma@unix.temple.edu Accepted 2006-DEC-8 by DAVET. 21-Dec-2006: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. J. Biol. Chem. (0021-9258) 2001 imex curation Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer lieberma@unix.temple.edu 21-Dec-2006: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Molecular weight was determined by fluorography. Molecular weight was determined by fluoragraphy. lieberma@unix.temple.edu Accepted 2006-DEC-8 by DAVET. 21-Dec-2006: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. J. Biol. Chem. (0021-9258) 2001 imex curation Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer lieberma@unix.temple.edu 21-Dec-2006: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Molecular weight was determined by fluorography. Possible request term weight by fluorography. Molecular weight was determined by fluorography. lieberma@unix.temple.edu Accepted 2006-DEC-8 by DAVET. 21-Dec-2006: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. J. Biol. Chem. (0021-9258) 2001 imex curation Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer lieberma@unix.temple.edu 21-Dec-2006: Contacted by JYOTI. 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 predetermined predetermined participant predetermined Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis lieberma@unix.temple.edu Accepted 2006-DEC-8 by DAVET. 21-Dec-2006: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. J. Biol. Chem. (0021-9258) 2001 imex curation Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer lieberma@unix.temple.edu 21-Dec-2006: Contacted by JYOTI. Only protein-protein interactions mouse-m1-1 Mus musculus m1 M1 human fibrobast murine myeloid cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Coimmunoprecipitation was carried out after IL-6 treatment. lieberma@unix.temple.edu Accepted 2006-DEC-8 by DAVET. 21-Dec-2006: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. Interaction of CR6 (GADD45gamma ) with proliferating cell nuclear antigen impedes negative growth control. J. Biol. Chem. (0021-9258) 2001 imex curation Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer lieberma@unix.temple.edu 21-Dec-2006: Contacted by JYOTI. Only protein-protein interactions 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 bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Azam N., Vairapandi M., Zhang W., Hoffman B., Liebermann DA. Apoptosis - Interactions involving proteins with a function related to apoptosis lieberma@unix.temple.edu Accepted 2006-DEC-8 by DAVET. 21-Dec-2006: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2001 Only protein-protein interactions imex curation pcna_human Proliferating cell nuclear antigen PCNA Cyclin protein protein human Homo sapiens Human MFEARLVQGSILKKVLEALKDLINEACWDISSSGVNLQSMDSSHVSLVQLTLRSEGFDTYRCDRNLAMGVNLTSMSKILKCAGNEDIITLRAEDNADTLALVFEAPNQEKVSDYEMKLMDLDVEQLGIPEQEYSCVVKMPSGEFARICRDLSHIGDAVVISCAKDGVKFSASGELGNGNIKLSQTSNVDKEEEAVTIEMNEPVQLTFALRYLNFFTKATPLSSTVTLSMSADVPLVVEYKIADMGHLKYYLAPKIEDEEGS DIP protein P12004 original sequence version: 200 DIP protein P12004 original sequence version: 128 DIP protein P12004 original sequence version: 139 DIP protein P12004 original sequence version: 185 DIP protein P12004 original sequence version: 102 DIP protein P12004 original sequence version: 105 DIP protein P12004 original sequence version: 158 E6F08E7EDBC48B00 ga45g_mouse Growth arrest and DNA damage-inducible protein GADD45 gamma Gadd45g Cr6 Cytokine-responsive protein CR6 protein protein mouse Mus musculus Mouse MTLEEVRGQDTVPESTARMQGAGKALHELLLSAHGQGCLTAGVYESAKVLNVDPDNVTFCVLAADEEDEGDIALQIHFTLIQAFCCENDIDIVRVGDVQRLAAIVGADEEGGAPGDLHCILISNPNEDTWKDPALEKLSLFCEESRSFNDWVPSITLPE DIP protein Q9Z111 original sequence version: 48 4B5996927C57988F cdn1a_mouse Cyclin-dependent kinase inhibitor 1 Cdkn1a Cip1 Waf1 p21 CDK-interacting protein 1 Melanoma differentiation-associated protein protein protein mouse Mus musculus Mouse MSNPGDVRPVPHRSKVCRCLFGPVDSEQLRRDCDALMAGCLQEARERWNFDFVTETPLEGNFVWERVRSLGLPKVYLSPGSRSRDDLGGDKRPSTSSALLQGPAPEDHVALSLSCTLVSERPEDSPGGPGTSQGRKRRQTSLTDFYHSKRRLVFCKRKP DIP protein P39689 original sequence version: 118 37B7C22B9A2FD089 pcna_mouse Proliferating cell nuclear antigen Pcna Cyclin protein protein mouse Mus musculus Mouse MFEARLIQGSILKKVLEALKDLINEACWDVSSGGVNLQSMDSSHVSLVQLTLRSEGFDTYRCDRNLAMGVNLTSMSKILKCAGNEDIITLRAEDNADTLALVFEAPNQEKVSDYEMKLMDLDVEQLGIPEQEYSCVIKMPSGEFARICRDLSHIGDAVVISCAKNGVKFSASGELGNGNIKLSQTSNVDKEEEAVTIEMNEPVHLTFALRYLNFFTKATPLSPTVTLSMSADVPLVVEYKIADMGHLKYYLAPKIEDEEAS DIP protein P17918 original sequence version: 136 DIP protein P17918 original sequence version: 111 DIP protein P17918 original sequence version: 129 F705CCBDD3205986 gadd45g-pcna 4 7 unspecified role unspecified role prey prey 8 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 1 A full-length CR6 interacted with PCNA pcna-gadd45g 3 7 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 8 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction 1 B, 4 A immune complexes of peptides corresponding to PCNA N-terminal (1-46 aa) and middle (87-127 aa) but not the C-terminal (224-261 aa) domains also contained CR6. Viewing figure 4 A, it seems that PCNA (87-127 aa) coimmunoprecipitates HA-CR6 but PCNA (1-46 aa) does not (unlike what is stated in the text), although PCNA (1-46 aa) is coimmunoprecipitated with HA-CR6. gadd45g-pcna-1 2 7 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 8 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 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 direct interaction direct interaction 1 B, 4 A immune complexes of peptides corresponding to PCNA N-terminal (1-46 aa) and middle (87-127 aa) but not the C-terminal (224-261 aa) domains also contained CR6. gadd45g-pcna-2 5 8 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 1 D the CR6 immune complex contained PCNA pcna-gadd45g-1 5 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association 1 D the PCNA immune complex, obtained from M1 cell extracts contained CR6. pcna-gadd45g-2 4 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 2 full-length PCNA interacted with CR6. PCNA C-terminal (PCNA/224�261), N-terminal (PCNA/1�46), and middle (PCNA/87�127) peptides also interacted with CR6. The interaction between full-length PCNA and full-length CR6 was weaker than the interaction when using PCNA/1-46 or PCNA/ 87-127 peptides. This observation is consistent with full-length PCNA containing in addition to CR6 interacting domain domains that hinder interaction. gadd45g-pcna-3 4 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait c-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 3 It can be seen that CR6/76�159 interacted with PCNA as well as full-length CR6, whereas CR6/1�135 failed to interact The binding affinity to PCNA of the CR6/76-159 C-terminal peptide was higher than the full-length CR6. pcna-gadd45g-3 3 8 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 region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 7 unspecified role unspecified role bait bait 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 direct interaction direct interaction 1 B, 4 B PCNA immune complexes contained the CR6/76-159 peptide gadd45g-pcna-4 2 8 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 7 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 direct interaction direct interaction 1 B, 4 B CR6 C-terminal (76-159 aa) immune complexes contained PCNA. gadd45g-pcna-5 1 7 unspecified role unspecified role prey prey 8 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 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 5 B HA immunocomplexes brought down PCNA from cells transfected with pcDNA3.1(2) encoding for C-terminal but not N-terminal CR6 pcna-gadd45g-4 6 7 unspecified role unspecified role bait bait 8 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 c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 5 B Transfection of 293T with pcDNA3.1(2) encoding for either C-terminal or N-terminal HA-CR6 peptides has demonstrated that PCNA immune complexes contained C-terminal but not N-terminal CR6 peptide. gadd45g-cdkn1a 5 9 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 1 D the CR6 immune complex contained p21. cdkn1a-gadd45g 5 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 1 D the p21 immune complex obtained from cell lysate after 48 h of IL-6 treatment contained CR6.