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/ Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Mol. Endocrinol. (0888-8809) 2004 imex curation Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. 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 anti tag western anti tag western blot anti tag western Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. Mol. Endocrinol. (0888-8809) 2004 jyoti Only protein-protein interactions imex curation Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Mol. Endocrinol. (0888-8809) 2004 imex curation Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp Accepted 2008-AUG-4 by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. Mol. Endocrinol. (0888-8809) 2004 jyoti Only protein-protein interactions imex curation Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Mol. Endocrinol. (0888-8809) 2004 imex curation Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. jyoti Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. Mol. Endocrinol. (0888-8809) 2004 jyoti Only protein-protein interactions imex curation Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Mol. Endocrinol. (0888-8809) 2004 imex curation Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. jyoti Only protein-protein interactions in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. Mol. Endocrinol. (0888-8809) 2004 jyoti Only protein-protein interactions imex curation Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Mol. Endocrinol. (0888-8809) 2004 imex curation Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. 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. 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 Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. Mol. Endocrinol. (0888-8809) 2004 jyoti Only protein-protein interactions imex curation Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Protein phosphatase 5 is a negative regulator of estrogen receptor-mediated transcription. Mol. Endocrinol. (0888-8809) 2004 imex curation Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. 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 Ikeda K., Ogawa S., Tsukui T., Horie-Inoue K., Ouchi Y., Kato S., Muramatsu M., Inoue S. Cancer - Interactions investigated in the context of cancer INOUE-GER@h.u-tokyo.ac.jp Accepted 2008-AUG-4 by JYOTI. 27-Aug-2008: Contacted by JYOTI. A cDNA library derived from estrogen-depleted MCF7 cells was used. Mol. Endocrinol. (0888-8809) 2004 jyoti Only protein-protein interactions imex curation esr1_human Estrogen receptor ESR1 ESR NR3A1 Estradiol receptor ER-alpha Nuclear receptor subfamily 3 group A member 1 protein protein human Homo sapiens Human MTMTLHTKASGMALLHQIQGNELEPLNRPQLKIPLERPLGEVYLDSSKPAVYNYPEGAAYEFNAAAAANAQVYGQTGLPYGPGSEAAAFGSNGLGGFPPLNSVSPSPLMLLHPPPQLSPFLQPHGQQVPYYLENEPSGYTVREAGPPAFYRPNSDNRRQGGRERLASTNDKGSMAMESAKETRYCAVCNDYASGYHYGVWSCEGCKAFFKRSIQGHNDYMCPATNQCTIDKNRRKSCQACRLRKCYEVGMMKGGIRKDRRGGRMLKHKRQRDDGEGRGEVGSAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATV DIP protein P03372 original sequence version: 214 DIP protein P03372 original sequence version: 139 DIP protein P03372 original sequence version: 167 5455C57AB0CCCAA7 ppp5_human Serine/threonine-protein phosphatase 5 Protein phosphatase T PPP5C PPP5 protein protein human Homo sapiens Human MAMAEGERTECAEPPRDEPPADGALKRAEELKTQANDYFKAKDYENAIKFYSQAIELNPSNAIYYGNRSLAYLRTECYGYALGDATRAIELDKKYIKGYYRRAASNMALGKFRAALRDYETVVKVKPHDKDAKMKYQECNKIVKQKAFERAIAGDEHKRSVVDSLDIESMTIEDEYSGPKLEDGKVTISFMKELMQWYKDQKKLHRKCAYQILVQVKEVLSKLSTLVETTLKETEKITVCGDTHGQFYDLLNIFELNGLPSETNPYIFNGDFVDRGSFSVEVILTLFGFKLLYPDHFHLLRGNHETDNMNQIYGFEGEVKAKYTAQMYELFSEVFEWLPLAQCINGKVLIMHGGLFSEDGVTLDDIRKIERNRQPPDSGPMCDLLWSDPQPQNGRSISKRGVSCQFGPDVTKAFLEENNLDYIIRSHEVKAEGYEVAHGGRCVTVFSAPNYCDQMGNKASYIHLQGSDLRPQFHQFTAVPHPNVKPMAYANTLLQLGMM DIP protein P53041 original sequence version: 76 DIP protein P53041 original sequence version: 168 DIP protein P53041 original sequence version: 169 DB3B2090D8658BB3 esr2_human Estrogen receptor beta ESR2 ESTRB NR3A2 Nuclear receptor subfamily 3 group A member 2 protein protein human Homo sapiens Human MDIKNSPSSLNSPSSYNCSQSILPLEHGSIYIPSSYVDSHHEYPAMTFYSPAVMNYSIPSNVTNLEGGPGRQTTSPNVLWPTPGHLSPLVVHRQLSHLYAEPQKSPWCEARSLEHTLPVNRETLKRKVSGNRCASPVTGPGSKRDAHFCAVCSDYASGYHYGVWSCEGCKAFFKRSIQGHNDYICPATNQCTIDKNRRKSCQACRLRKCYEVGMVKCGSRRERCGYRLVRRQRSADEQLHCAGKAKRSGGHAPRVRELLLDALSPEQLVLTLLEAEPPHVLISRPSAPFTEASMMMSLTKLADKELVHMISWAKKIPGFVELSLFDQVRLLESCWMEVLMMGLMWRSIDHPGKLIFAPDLVLDRDEGKCVEGILEIFDMLLATTSRFRELKLQHKEYLCVKAMILLNSSMYPLVTATQDADSSRKLAHLLNAVTDALVWVIAKSGISSQQQSMRLANLLMLLSHVRHASNKGMEHLLNMKCKNVVPVYDLLLEMLNAHVLRGCKSSITGSECSPAEDSKSKEGSQNPQSQ DIP protein Q92731 original sequence version: 171 8CAE34215992418A esr2-ppp5c 2 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 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 physical association physical association The Estrogen receptor (ER) beta construct aa 1-481 is a dominant negative form, that has the potential to repress both ERalpha- and ERbeta-mediated transactivation. A yeast two-hybrid screening identified a serine/threonine protein phosphatase (PP5) as an interactant of ER (1-481), a dominant negative ER mutant. esr2-ppp5c-1 6 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 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 physical association physical association The Estrogen receptor (ER) beta construct aa 1-481 is a dominant negative form, that has the potential to repress both ERalpha- and ERbeta-mediated transactivation. The interactions of the PP5 clones between ER (1-481) were confirmed by the galactose-dependent growth of yeast strain EGY48, which was cotransformed with pSH18-34 LacZ reporter plasmid, pEG202NLS-ER (1-481), and the PP5 clones in galactose-inducible pJG4-5 1 ppp5c-esr2 4 8 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 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 physical association physical association 2 This interaction is not ligand-dependent. The level of interaction was not affected by the presence or absence of E2. Both ER and ER were pulled down by full-length PP5 ppp5c-esr1 4 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 8 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association 2 This interaction is not ligand-dependent. The level of interaction was not affected by the presence or absence of E2. Both ER and ER were pulled down by full-length PP5 esr2-ppp5c-2 5 9 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 8 unspecified role unspecified role prey prey physical association physical association 3 The most significant luciferase activity was observed when PP5 interacted with E domains of ERs without containing the activation domain (AD) core regions within AF-2 The highest level of interaction was observed when the ERbeta construct containing only the E domain (aa 248-481) was used. esr1-ppp5c 5 8 unspecified role unspecified role prey prey 7 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 physical association physical association 3 The most significant luciferase activity was observed when PP5 interacted with E domains of ERs without containing the activation domain (AD) core regions within AF-2 The highest levels of this interaction were observed when the ERalpha construct containing only the E domain (aa 302-530) was used. ppp5c-esr1-1 1 8 unspecified role unspecified role bait bait 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 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 region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 4 B The finding that TPR domains in PP5 are requisite sites for interaction with ERs was further confirmed by coimmunoprecipitation experiments, using 293T cells cotransfected with green fluorescent protein (GFP)-tagged ER along with Flag-tagged full-length PP5 or a PP5 mutant with only its TPR domains ppp5c-esr1-2 3 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 4 C The in vivo interaction between endogenous PP5 and ER in human breast cancer MCF7 cells was also shown by coimmunoprecipitation study