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/ http://www.ebi.ac.uk/ stambolsky-2010-1 Modulation of the vitamin D3 response by cancer-associated mutant p53. human-skbr3 Homo sapiens SKBR3 skbr3 SKBR3 Species: human; Tissue: breast; Tumor: adenocarcinoma; Derived from: metastatic site: pleural effusion chip-chip chromatin immunoprecipitation array chip-chip nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Stambolsky P., Tabach Y., Fontemaggi G., Weisz L., Maor-Aloni R., Sigfried Z., Shiff I., Kogan I., Shay M., Kalo E., Blandino G., Simon I., Oren M., Rotter V. Apoptosis - Interactions involving proteins with a function related to apoptosis Hu13K human promoter array was used. perrystam@hotmail.com Cancer Cell (1535-6108) 2010 Only protein-protein interactions 5 imex curation Cancer - Interactions investigated in the context of cancer stambolsky-2010-6 Modulation of the vitamin D3 response by cancer-associated mutant p53. human-skbr3 Homo sapiens SKBR3 skbr3 SKBR3 Species: human; Tissue: breast; Tumor: adenocarcinoma; Derived from: metastatic site: pleural effusion ch-ip chromatin immunoprecipitation assay ch-ip nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Stambolsky P., Tabach Y., Fontemaggi G., Weisz L., Maor-Aloni R., Sigfried Z., Shiff I., Kogan I., Shay M., Kalo E., Blandino G., Simon I., Oren M., Rotter V. Apoptosis - Interactions involving proteins with a function related to apoptosis Hu13K human promoter array was used. perrystam@hotmail.com Cancer Cell (1535-6108) 2010 Only protein-protein interactions 5 imex curation Cancer - Interactions investigated in the context of cancer stambolsky-2010-2 Modulation of the vitamin D3 response by cancer-associated mutant p53. human-skbr3 Homo sapiens SKBR3 skbr3 SKBR3 Species: human; Tissue: breast; Tumor: adenocarcinoma; Derived from: metastatic site: pleural effusion filter binding filter binding Filter overlay assay Filter overlay assay western blot western blot Immuno blot Immuno blot Stambolsky P., Tabach Y., Fontemaggi G., Weisz L., Maor-Aloni R., Sigfried Z., Shiff I., Kogan I., Shay M., Kalo E., Blandino G., Simon I., Oren M., Rotter V. Apoptosis - Interactions involving proteins with a function related to apoptosis Hu13K human promoter array was used. perrystam@hotmail.com Cancer Cell (1535-6108) 2010 Only protein-protein interactions 5 imex curation Cancer - Interactions investigated in the context of cancer stambolsky-2010-4 Modulation of the vitamin D3 response by cancer-associated mutant p53. human-skbr3 Homo sapiens SKBR3 skbr3 SKBR3 Species: human; Tissue: breast; Tumor: adenocarcinoma; Derived from: metastatic site: pleural effusion anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Stambolsky P., Tabach Y., Fontemaggi G., Weisz L., Maor-Aloni R., Sigfried Z., Shiff I., Kogan I., Shay M., Kalo E., Blandino G., Simon I., Oren M., Rotter V. Apoptosis - Interactions involving proteins with a function related to apoptosis Hu13K human promoter array was used. perrystam@hotmail.com Cancer Cell (1535-6108) 2010 Only protein-protein interactions 5 imex curation Cancer - Interactions investigated in the context of cancer stambolsky-2010-5 Modulation of the vitamin D3 response by cancer-associated mutant p53. 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 Stambolsky P., Tabach Y., Fontemaggi G., Weisz L., Maor-Aloni R., Sigfried Z., Shiff I., Kogan I., Shay M., Kalo E., Blandino G., Simon I., Oren M., Rotter V. Apoptosis - Interactions involving proteins with a function related to apoptosis Hu13K human promoter array was used. perrystam@hotmail.com Cancer Cell (1535-6108) 2010 Only protein-protein interactions 5 imex curation Cancer - Interactions investigated in the context of cancer stambolsky-2010-8 Modulation of the vitamin D3 response by cancer-associated mutant p53. human-mda_mb_231 Homo sapiens Caucasian breast adenocarcinoma mda_mb_231 Human Caucasian breast adenocarcinoma ch-ip chromatin immunoprecipitation assay ch-ip nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Stambolsky P., Tabach Y., Fontemaggi G., Weisz L., Maor-Aloni R., Sigfried Z., Shiff I., Kogan I., Shay M., Kalo E., Blandino G., Simon I., Oren M., Rotter V. Apoptosis - Interactions involving proteins with a function related to apoptosis Hu13K human promoter array was used. perrystam@hotmail.com Cancer Cell (1535-6108) 2010 Only protein-protein interactions 5 imex curation Cancer - Interactions investigated in the context of cancer stambolsky-2010-3 Modulation of the vitamin D3 response by cancer-associated mutant p53. human-h1299 Homo sapiens lung lymph node carcinoma h1299 Human lung lymph node carcinoma p53 null. These cells have also been reported to secrete the peptide hormone neuromedin B (NMB), but not gastrin releasing peptide (GRP). Also known as NCI H1299 or CRL-5803. http://www.lgcpromochem.com/atcc/ ATCC number: CRL-5803. ATCC number of the cell line is CRL-5803. fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Stambolsky P., Tabach Y., Fontemaggi G., Weisz L., Maor-Aloni R., Sigfried Z., Shiff I., Kogan I., Shay M., Kalo E., Blandino G., Simon I., Oren M., Rotter V. Apoptosis - Interactions involving proteins with a function related to apoptosis Hu13K human promoter array was used. perrystam@hotmail.com Cancer Cell (1535-6108) 2010 Only protein-protein interactions 5 imex curation Cancer - Interactions investigated in the context of cancer vipr2_human_gene Human VIPR2 promoter gene gene human Homo sapiens Human p53_human Cellular tumor antigen p53 TP53 P53 Tumor suppressor p53 Phosphoprotein p53 Antigen NY-CO-13 protein protein human Homo sapiens Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD mkl2_human_gene Human MKL2 gene gene gene human Homo sapiens Human mageh1_human_gene Human MAGEH1 promoter gene gene human Homo sapiens Human ece2_human_gene Human ECE2 promoter gene gene human Homo sapiens Human fras1_human_gene Human FRAS1 promoter gene gene human Homo sapiens Human ghr_human_gene Human Growth Hormone Receptor Promoter gene gene human Homo sapiens Human mark2_human_gene Human MARK2 promoter gene gene human Homo sapiens Human cpt1b_human _gene Human CPT1B (muscle isoform) promoter gene gene human Homo sapiens Human SwissProt entry is Q92523 skp2_human_gene Human SKP2 promoter gene gene human Homo sapiens Human nfs1_human_gene Human NFS1 gene gene gene human Homo sapiens Human ecrg4_human_gene Human ECRG4 promoter gene gene human Homo sapiens Human klf10_human_gene Human KLF10 promoter gene gene human Homo sapiens Human actr10_human_gene Human ACTR10 promoter gene gene human Homo sapiens Human lig1_human_gene Human LIG1 promoter gene gene human Homo sapiens Human stath_human_gene Human STATH promoter gene gene human Homo sapiens Human tuba1c_human_gene Human TUBA1C promoter gene gene human Homo sapiens Human c12orf49_human_gene Human C12orf49 promoter gene gene human Homo sapiens Human pcdha7_human_gene Human PCDHA7 promoter gene gene human Homo sapiens Human hira_human_gene Human HIRA promoter VDRE (Vitamin D Response Element) gene gene human Homo sapiens Human vdr_human Vitamin D3 receptor 1,25-dihydroxyvitamin D3 receptor Nuclear receptor subfamily 1 group I member 1 VDR NR1I1 protein protein human Homo sapiens Human MEAMAASTSLPDPGDFDRNVPRICGVCGDRATGFHFNAMTCEGCKGFFRRSMKRKALFTCPFNGDCRITKDNRRHCQACRLKRCVDIGMMKEFILTDEEVQRKREMILKRKEEEALKDSLRPKLSEEQQRIIAILLDAHHKTYDPTYSDFCQFRPPVRVNDGGGSHPSRPNSRHTPSFSGDSSSSCSDHCITSSDMMDSSSFSNLDLSEEDSDDPSVTLELSQLSMLPHLADLVSYSIQKVIGFAKMIPGFRDLTSEDQIVLLKSSAIEVIMLRSNESFTMDDMSWTCGNQDYKYRVSDVTKAGHSLELIEPLIKFQVGLKKLNLHEEEHVLLMAICIVSPDRPGVQDAALIEAIQDRLSNTLQTYIRCRHPPPGSHLLYAKMIQKLADLRSLNEEHSKQYRCLSFQPECSMKLTPLVLEVFGNEIS tgfb1_human_gene TGFB1_HUMAN_GENE gene gene human Homo sapiens Human cp24a_human_gene Human CYP24A1 promoter VDRE (Vitamin D Response Element) gene gene human Homo sapiens Human cite1_human Cbp/p300-interacting transactivator 1 CITED1 MSG1 Melanocyte-specific protein 1 protein protein human Homo sapiens Human MPTTSRPALDVKGGTSPAKEDANQEMSSVAYSNLAVKDRKAVAILHYPGVASNGTKASGAPTSSSGSPIGSPTTTPPTKPPSFNLHPAPHLLASMHLQKLNSQYQGMAAATPGQPGEAGPLQNWDFGAQAGGAESLSPSAGAQSPAIIDSDPVDEEVLMSLVVELGLDRANELPELWLGQNEFDFTADFPSSC cp27b_human_gene Human CYP27B1 promoter nVDRE (negative Vitamin D Response Element) gene gene human Homo sapiens Human vdre_probe_human Human Vitamin D Response Element VDRE: consensus oligonucleotide dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid human Homo sapiens Human AGGTCANNNAGGTCA VDRE consensus oligonucleotide sequence. mutp53-vipr2 894230 n/a 894237 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-mkl2 894230 n/a 894239 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-mageh1 894230 n/a 894240 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-ece2 894230 n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 894241 unspecified role unspecified role prey prey association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-fras1 894230 n/a 894242 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-ghr 894230 n/a 894243 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-mark2 894230 n/a 894244 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-cpt1b 894230 n/a 894245 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-skp2 894230 n/a 894246 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-nfs1 894230 n/a 894247 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-ecrg4 894230 n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 894248 unspecified role unspecified role prey prey association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-klf10 894230 n/a 894249 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-actr10 894230 n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 894250 unspecified role unspecified role prey prey association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-lig1 894230 n/a 894251 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-stath 894230 n/a 894252 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-tuba1c 894230 n/a 894253 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-c12orf49 894230 n/a 894254 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). mutp53-pcdha7 894230 n/a 894255 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Table 1 One of thirty Gene Promoters preferentially Bound by Mutp53(R175H). vdr-hira 894231 n/a 894256 unspecified role unspecified role prey prey n/a 894257 unspecified role unspecified role bait bait association association false false false Fig 1A. Vitamin D3 Significant binding of mutp53 to the VDRE-containing regions of both HIRA and TGF-B1 promoters was clearly detectable, and Vitamin D3 treatment markedly increased binding. p53mut-hira 894231 n/a 894256 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Fig 1A. Vitamin D3 Significant binding of mutp53 to the VDRE-containing regions of both HIRA and TGF-B1 promoters was clearly detectable, and Vitamin D3 treatment markedly increased binding. p53mut-tgfb1 894231 n/a 894258 unspecified role unspecified role prey prey promoter vdre (vitamin d response element) binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Fig 1A. Vitamin D3 Significant binding of mutp53 to the VDRE-containing regions of both HIRA and TGF-B1 promoters was clearly detectable, and Vitamin D3 treatment markedly increased binding. vdr-tgfb1 894231 n/a 894258 unspecified role unspecified role prey prey promoter vdre (vitamin d response element) binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 894257 unspecified role unspecified role bait bait association association false false false Fig 1A. Vitamin D3 Significant binding of mutp53 to the VDRE-containing regions of both HIRA and TGF-B1 promoters was clearly detectable, and Vitamin D3 treatment markedly increased binding. vdr-cp24a 894231 n/a 894259 unspecified role unspecified role prey prey n/a 894257 unspecified role unspecified role bait bait association association false false false Fig 3D. Vitamin D3 mutp53, VDR, and p300 bound to the CYP24A1 promoter VDRE. p53mut-cyp24a 894231 n/a 894259 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false association association false false false Fig 3D. Vitamin D3 mutp53, VDR, and p300 bound to the CYP24A1 promoter VDRE. p300-cyp24a-1 894231 n/a 894260 unspecified role unspecified role bait bait n/a 894259 unspecified role unspecified role prey prey association association false false false Fig 3D. Vitamin D3 mutp53, VDR, and p300 bound to the CYP24A1 promoter VDRE. cite1-cp27b 894231 n/a 894261 unspecified role unspecified role prey prey n/a 894260 unspecified role unspecified role bait bait association association false false false Fig 3G. Vitamin D3 Mutp53 was not associated with the region of the CYP27B gene harboring the nVDRE. vdre-mutp53 894232 n/a 894262 unspecified role unspecified role bait bait n/a 894238 unspecified role unspecified role prey prey P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig S1B. mutp53 displayed weak specific binding to the VDRE under basal conditions, which was significantly augmented upon vitamin D3 treatment. 100nM vitamin D3 vdre-vdr 894232 n/a 894262 unspecified role unspecified role bait bait n/a 894257 unspecified role unspecified role prey prey physical association physical association false false false Fig S1B. Vitamin D3 treatment augmented the binding of VDR to a wild-type but not mutant VDRE sequence, as expected. 100nM vitamin D3 vdr-mutp53-1 894233 n/a 894257 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig 3A. Vitamin D3 Reciprocal coimmunoprecipitations carried out with either a mutp53-specific antibody or anti-VDR antibody. vdr-mutp53-2 894233 n/a 894238 unspecified role unspecified role prey prey P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 894257 unspecified role unspecified role bait bait physical association physical association false false false Fig 3A, Fig S2A. Vitamin D3 Pull-down of VDR coprecipitated p53 from both SKBR3 and MCF7 cell extracts. The coimmunoprecipitation of mutp53 with VDR was further augmented when a wild-type, but not mutant, VDRE consensus oligonucleotide was added to the cell extract during incubation, indicating that the VDRE sequence can recruit both proteins simultaneously. Wild type VDRE consensus oligonucleotide vdr-mutp53-3 894234 n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 894257 unspecified role unspecified role prey prey physical association physical association false false false Fig 3A. Vitamin D3 Reciprocal coimmunoprecipitations carried out with either a mutp53-specific antibody or anti-VDR antibody. vdr-mutp53-4 894234 n/a 894238 unspecified role unspecified role prey prey n/a 894257 unspecified role unspecified role bait bait physical association physical association false false false Fig 3A. Vitamin D3 Pull-down of VDR coprecipitated p53 from both SKBR3 and MCF7 cell extracts. vdr-cp24a-p53 894235 n/a 894257 unspecified role unspecified role bait bait n/a 894238 unspecified role unspecified role prey prey P04637:p.Arg280Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 894259 unspecified role unspecified role prey prey association association false false false Fig 3E. Vitamin D3 ChIP for p53 followed by re-ChIP for VDR or p300 enriched for the presence of the CYP24A VDRE when treated with vitamin D3. p300-cyp24a-p53 894235 n/a 894238 unspecified role unspecified role prey prey P04637:p.Arg280Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false n/a 894259 unspecified role unspecified role prey prey n/a 894260 unspecified role unspecified role bait bait association association false false false Fig 3E. Vitamin D3 ChIP for p53 followed by re-ChIP for VDR or p300 enriched for the presence of the CYP24A VDRE when treated with vitamin D3. vdr-mutp53 894236 n/a 894257 unspecified role unspecified role prey prey n/a 894238 unspecified role unspecified role bait bait P04637:p.Arg175His mutation mutation certain certain sequence position certain certain certain sequence position certain false colocalization colocalization false false false Fig 2D. Zn2+ (to induce p53 expression). Vitamin D3. In untreated H1299R175H-i cells VDR is largely cytoplasmic. Combining mutp53 induction with vitamin D3 rendered VDR staining more prominent and almost exclusively nuclear.