Evidence for different modes of action of cyclin-dependent kinase inhibitors: p15 and p16 bind to kinases, p21 and p27 bind to cyclins.
A number of low molecular weight proteins have recently been identified that specifically inhibit the function of cyclin-dependent protein kinases in mammalian cells. These fall into two distinct families based on primary sequence comparisons and probable modes of action. Using a simple in vitro binding assay, we show that p21CDKN1 and the related p27KIP1 can efficiently interact with cyclins D1, D2, D3, E and A, and to a lesser extent with cyclin B. By generating a deleted form of cyclin D1 that binds to p21 and p27 but not to Cdks, we confirm that these interaction do not depend on stoichiometric amounts of the relevant kinase subunit. Moreover, p21 and p27 do not detectably associated with kinase subunits unless a cyclin is present. This is in sharp contrast to the properties of p16CDKN2 and p15MTS2/INK4b which bind to Cdk4 and Cdk6 in the absence of any cyclin. These data suggest that p21 and p27 act as broad spectrum regulators of cyclin dependent kinase function by participating in ternary complexes whereas the p16 family specifically interfere with the formation of cyclin D-dependent kinase complexes.
D002352:Carrier Proteins / Q000378:metabolism*; D018797:Cell Cycle Proteins; D050762:Cyclin-Dependent Kinase Inhibitor p15; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050759:Cyclin-Dependent Kinase Inhibitor p21; D050760:Cyclin-Dependent Kinase Inhibitor p27; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D016213:Cyclins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D006801:Humans; D007158:Immunologic Techniques; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D011485:Protein Binding; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins
C493956:CDKN1A protein, human; C493966:CDKN2B protein, human; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D050762:Cyclin-Dependent Kinase Inhibitor p15; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D008869:Microtubule-Associated Proteins; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D018844:Cyclin-Dependent Kinases