Cyclin D1/cdk2 kinase is present in a G1 phase-specific protein complex Yi1 that binds to the mouse thymidine kinase gene promoter.
Our studies provide evidence for the presence of cyclin D1 in an early G1 cycle-specific DNA binding complex Yi1. Previously we identified several complexes including Yi and E2F that at different times during G0 to S transition bind to three distinct DNA sequences (MT1, MT2, MT3) located in the mouse thymidine kinase upstream promoter. These various complexes contain DNA binding proteins (Sp1, E2F, p110, p60), cyclins A and E, cyclin-dependent kinase 2 (cdk2), and retinoblastoma-related proteins (pRB, p107). Here we report that Yi1 is different from the E2F complexes. Yi1 contains cyclin D1/cdk2 kinase as shown by using specific antibodies to cyclins, cdks and the Yi1 DNA-binding protein in gel retardation, western blotting, and immunoprecipitation assays. Yi1 binding is specific to a consensus sequence different from that of E2F.
Dou
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Department of Pharmacology, University of Pittsburgh School of Medicine, PA.
Molnar
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Pardee
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D016475:3T3 Cells; D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D042846:CDC2-CDC28 Kinases; D019938:Cyclin D1; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D016213:Cyclins / Q000276:immunology / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D016193:G1 Phase; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D015513:Oncogene Proteins / Q000276:immunology / Q000378:metabolism*; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D013937:Thymidine Kinase / Q000235:genetics*
D000906:Antibodies; D016213:Cyclins; D046911:Macromolecular Substances; D015513:Oncogene Proteins; D019938:Cyclin D1; D004247:DNA; D013937:Thymidine Kinase; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495893:Cdk2 protein, mouse; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases