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 CDK10 as an Important Determinant of Resistance to Endocrine Therapy for Breast Cancer. Identification of CDK10 as an Important Determinant of Resistance to Endocrine Therapy for Breast Cancer. Cancer Cell (1535-6108) 2008 imex curation Iorns E., Turner NC., Elliott R., Syed N., Garrone O., Gasco M., Tutt AN., Crook T., Lord CJ., Ashworth A. Cancer - Interactions investigated in the context of cancer The product number provided by the authors was used to search the Genecopoeia website, which shows the CDK10 construct used to be of human origin. chris.lord@icr.ac.uk,alan.ashworth@icr.ac.uk 14-May-2008: Contacted by JYOTI. davet Only protein-protein interactions 2 human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells ch-ip chromatin immunoprecipitation assay ch-ip primer specific pcr primer specific pcr Iorns E., Turner NC., Elliott R., Syed N., Garrone O., Gasco M., Tutt AN., Crook T., Lord CJ., Ashworth A. Cancer - Interactions investigated in the context of cancer The product number provided by the authors was used to search the Genecopoeia website, which shows the CDK10 construct used to be of human origin. chris.lord@icr.ac.uk,alan.ashworth@icr.ac.uk Accepted 2008-APR-19 by DAVET. 14-May-2008: Contacted by JYOTI. Cancer Cell (1535-6108) 2008 davet Only protein-protein interactions 2 imex curation Identification of CDK10 as an Important Determinant of Resistance to Endocrine Therapy for Breast Cancer. Identification of CDK10 as an Important Determinant of Resistance to Endocrine Therapy for Breast Cancer. Cancer Cell (1535-6108) 2008 imex curation Iorns E., Turner NC., Elliott R., Syed N., Garrone O., Gasco M., Tutt AN., Crook T., Lord CJ., Ashworth A. Cancer - Interactions investigated in the context of cancer The product number provided by the authors was used to search the Genecopoeia website, which shows the CDK10 construct used to be of human origin. chris.lord@icr.ac.uk,alan.ashworth@icr.ac.uk 14-May-2008: Contacted by JYOTI. davet Only protein-protein interactions 2 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 Iorns E., Turner NC., Elliott R., Syed N., Garrone O., Gasco M., Tutt AN., Crook T., Lord CJ., Ashworth A. Cancer - Interactions investigated in the context of cancer The product number provided by the authors was used to search the Genecopoeia website, which shows the CDK10 construct used to be of human origin. chris.lord@icr.ac.uk,alan.ashworth@icr.ac.uk Accepted 2008-APR-19 by DAVET. 14-May-2008: Contacted by JYOTI. Cancer Cell (1535-6108) 2008 davet Only protein-protein interactions 2 imex curation Identification of CDK10 as an Important Determinant of Resistance to Endocrine Therapy for Breast Cancer. Identification of CDK10 as an Important Determinant of Resistance to Endocrine Therapy for Breast Cancer. Cancer Cell (1535-6108) 2008 imex curation Iorns E., Turner NC., Elliott R., Syed N., Garrone O., Gasco M., Tutt AN., Crook T., Lord CJ., Ashworth A. Cancer - Interactions investigated in the context of cancer The product number provided by the authors was used to search the Genecopoeia website, which shows the CDK10 construct used to be of human origin. chris.lord@icr.ac.uk,alan.ashworth@icr.ac.uk 14-May-2008: Contacted by JYOTI. davet Only protein-protein interactions 2 human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Iorns E., Turner NC., Elliott R., Syed N., Garrone O., Gasco M., Tutt AN., Crook T., Lord CJ., Ashworth A. Cancer - Interactions investigated in the context of cancer The product number provided by the authors was used to search the Genecopoeia website, which shows the CDK10 construct used to be of human origin. chris.lord@icr.ac.uk,alan.ashworth@icr.ac.uk Accepted 2008-APR-19 by DAVET. 14-May-2008: Contacted by JYOTI. Cancer Cell (1535-6108) 2008 davet Only protein-protein interactions 2 imex curation raf1_human_gene Human RAF1 (c-RAF) gene gene gene human Homo sapiens Human cdk10_human Cyclin-dependent kinase 10 CDK10 Serine/threonine-protein kinase PISSLRE Cell division protein kinase 10 protein protein human Homo sapiens Human MAEPDLECEQIRLKCIRKEGFFTVPPEHRLGRCRSVKEFEKLNRIGEGTYGIVYRARDTQTDEIVALKKVRMDKEKDGIPISSLREITLLLRLRHPNIVELKEVVVGNHLESIFLVMGYCEQDLASLLENMPTPFSEAQVKCIVLQVLRGLQYLHRNFIIHRDLKVSNLLMTDKGCVKTADFGLARAYGVPVKPMTPKVVTLWYRAPELLLGTTTQTTSIDMWAVGCILAELLAHRPLLPGTSEIHQIDLIVQLLGTPSENIWPGFSKLPLVGQYSLRKQPYNNLKHKFPWLSEAGLRLLHFLFMYDPKKRATAGDCLESSYFKEKPLPCEPELMPTFPHHRNKRAAPATSEGQSKRCKP 25B28560961103F4 ets2_human Protein C-ets-2 ETS2 protein protein human Homo sapiens Human MNDFGIKNMDQVAPVANSYRGTLKRQPAFDTFDGSLFAVFPSLNEEQTLQEVPTGLDSISHDSANCELPLLTPCSKAVMSQALKATFSGFKKEQRRLGIPKNPWLWSEQQVCQWLLWATNEFSLVNVNLQRFGMNGQMLCNLGKERFLELAPDFVGDILWEHLEQMIKENQEKTEDQYEENSHLTSVPHWINSNTLGFGTEQAPYGMQTQNYPKGGLLDSMCPASTPSVLSSEQEFQMFPKSRLSSVSVTYCSVSQDFPGSNLNLLTNNSGTPKDHDSPENGADSFESSDSLLQSWNSQSSLLDVQRVPSFESFEDDCSQSLCLNKPTMSFKDYIQERSDPVEQGKPVIPAAVLAGFTGSGPIQLWQFLLELLSDKSCQSFISWTGDGWEFKLADPDEVARRWGKRKNKPKMNYEKLSRGLRYYYDKNIIHKTSGKRYVYRFVCDLQNLLGFTPEELHAILGVQPDTED DIP protein P15036 original sequence version: 158 5944EC4B5AAB553E cdk10-ets2 3 5 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 6 unspecified role unspecified role prey prey physical association physical association 7 D We confirmed that CDK10 binds to ETS2 using coimmunoprecipitation Q15131 transfection ets2-cdk10 2 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association S5 We confirmed that CDK10 binds to ETS2 using coimmunoprecipitation cdk10-raf1_human 1 5 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 4 unspecified role unspecified role prey prey association association 7 F identified a putative ETS2-binding site in the c-RAF promoter (Figure 7E). We demonstrated using chromatin IP (ChIP) that ETS2 and CDK10 bind to this site Q15131 transfection CDK10 binds to the ETS2-binding site in the c-RAF promoter (GGAT). ets2-raf1_human-1 1 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait association association 7 F, G identified a putative ETS2-binding site in the c-RAF promoter (Figure 7E). We demonstrated using chromatin IP (ChIP) that ETS2 and CDK10 bind to this site Q15131 transfection Silencing of CDK10 leads to enhanced binding of ETS2 to the ETS2 binding site in the c-RAF promoter. CDK10 - Transfection with the CDK10 expression vector decreases this interaction. ETS2 binds to the ETS2-binding site in the c-RAF promoter (GGAT).