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/ Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. J. Biol. Chem. (0021-9258) 2007 Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. orchard human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk Accepted 2007-Not a month!-5 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. J. Biol. Chem. (0021-9258) 2007 orchard Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. J. Biol. Chem. (0021-9258) 2007 Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. orchard human-hpr_1 Homo sapiens normal prostate epithelial cell line hpr_1 Human normal prostate epithelial cell line fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk Accepted 2007-Not a month!-5 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. J. Biol. Chem. (0021-9258) 2007 orchard Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. J. Biol. Chem. (0021-9258) 2007 Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. orchard human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk Accepted 2007-Not a month!-5 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. J. Biol. Chem. (0021-9258) 2007 orchard Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. J. Biol. Chem. (0021-9258) 2007 Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. orchard human-du_145 Homo sapiens DU-145 prostate cancer cell line du_145 DU-145 Human prostate carcinoma established from the tumor tissue removed from the metastatic central nervous system lesion of a 69-year-old man with prostate carcinoma in 1975 confirmed as human with IEF of AST, MDH, NP anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk Accepted 2007-Not a month!-5 by ORCHARD. 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. J. Biol. Chem. (0021-9258) 2007 orchard Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. Identification of a Novel Inhibitor of Differentiation-1 (ID-1) Binding Partner, Caveolin-1, and Its Role in Epithelial-Mesenchymal Transition and Resistance to Apoptosis in Prostate Cancer Cells. J. Biol. Chem. (0021-9258) 2007 Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk 22-Jan-2008: Contacted by JYOTI. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. orchard 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 Zhang X., Ling MT., Wang Q., Lau CK., Leung SC., Lee TK., Cheung AL., Wong YC., Wang X. Apoptosis - Interactions involving proteins with a function related to apoptosis PMID 12016143 with the last author as one of the authors describes the Id-1 construct as of human origin and a number of papers by Wang, X. refer back to this reference as describing the Id-1 construct. ycwong@hkucc.hku.hk Accepted 2007-Not a month!-5 by ORCHARD. A cDNA library derived from a human nonmalignant prostatic epithelial cell line, Hpr-1, was used. J. Biol. Chem. (0021-9258) 2007 orchard id1_human DNA-binding protein inhibitor ID-1 ID1 ID Inhibitor of DNA binding 1 BHLHB24 Class B basic helix-loop-helix protein 24 Inhibitor of differentiation 1 protein protein human Homo sapiens Human MKVASGSTATAAAGPSCALKAGKTASGAGEVVRCLSEQSVAISRCAGGAGARLPALLDEQQVNVLLYDMNGCYSRLKELVPTLPQNRKVSKVEILQHVIDYIRDLQLELNSESEVGTPGGRGLPVRAPLSTLNGEISALTAEAACVPADDRILCR DIP protein P41134 original sequence version: 156 DIP protein P41134 original sequence version: 115 480287384B667161 cav1_human Caveolin-1 CAV1 CAV protein protein human Homo sapiens Human MSGGKYVDSEGHLYTVPIREQGNIYKPNNKAMADELSEKQVYDAHTKEIDLVNRDPKHLNDDVVKIDFEDVIAEPEGTHSFDGIWKASFTTFTVTKYWFYRLLSALFGIPMALIWGIYFAILSFLHIWAVVPCIKSFLIEIQCISRVYSIYVHTVCDPLFEAVGKIFSNVRINLQKEI DIP protein Q03135 original sequence version: 164 DIP protein Q03135 original sequence version: 119 DIP protein Q03135 original sequence version: 122 DIP protein Q03135-1 original sequence version: 122 DIP protein Q03135 original sequence version: 133 DIP protein Q03135 original sequence version: 155 DIP protein Q03135 original sequence version: 147 2424DE9B5E6521D5 id1-cav1 5 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association To identify novel Id-1 interacting proteins, we used a full-length Id-1 cDNA as a bait to screen a cDNA library derived from an immortalized human nonmalignant prostatic epithelial cell line, Hpr-1, using the yeast two-hybrid technique. Sixteen positive clones were selected (data not shown). Sequence analysis and data base comparison revealed that one positive clone had 100% identity to the Homo sapiens caveolin-1 mRNA coding sequence PREY gene NM_001753 id1-cav1-1 1 6 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 required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain P41134:p.Tyr101Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y D Wed May 02 18:13:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr101asp' Tue Jun 30 17:31:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr101Asp' 7 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 1 A, E After immunoprecipitation with anti-FLAG M2 affinity resin, Cav-1-HA was detected in the precipitates derived from the cells co-transfected with both Id-1-FLAG and Cav-1-HA (Fig. 1A, lane 4), indicating co-immunoprecipitation of Cav-1 with Id-1. P41134; Q03135 transfection PREY gene NM_001753 The deletion analysis and mutational analysis suggest that the HLH domain (aa 66-106) of Id-1 is essential for its interaction with Cav-1. cav1-id1 1 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 7 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 Reciprocally, after immunoprecipitation with anti-HA-agarose affinity resin (Fig. 1A, lower panel), Id-1-FLAG was detected in the cells co-transfected with the two proteins (lane 8), confirming the interaction between Cav-1 and Id-1. Q03135; P41134 transfection BAIT gene NM_001753 cav1-id1-1 4 7 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association 1 B endogenous Id-1 was detected in the immunoprecipitates by an anti-Cav-1 antibody cav1-id1-2 3 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association 1 B endogenous Id-1 was detected in the immunoprecipitates by an anti-Cav-1 antibody cav1-id1-3 2 7 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component colocalization colocalization 1 C fluorescent immunostaining also showed that both Id-1 and Cav-1 were co-localized in the cytoplasm of Hpr-1 cells