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Involvement of p27 in myc-regulated mammalian cell proliferation

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The cell proliferation regulatory mechanism by Myc remains unclear. In a model of rat fibroblasts transfected by MycER conditional chimeras, activation of Myc induces the increase of cyclin E/cdk2
complexes-associated catalytic activity. Further data argued for an involvement of the cdk inhibitor p27 in this process since Myc activation of cyclin E-complexes correlated with the dissociation of p27 from these complexes and its loss from whole-cell lysates. The objectives of the present research proposal are 1) to provide some direct evidence that p27 is a relevant target for Myc and clarify its precise role in Myc biology. For this purpose, the effects of Myc activation on cyclin/cdk kinase activity will be analyzed by biochemical experiments in p27 knockout mice. 2) to identify the components induced by Myc, involved in degradation or in sequestration of p27. To address this issue, a genetic approach using the two hybrid system to screen in a rat fibroblast library the p27-interacting proteins complemented by a biochemical approach using gel exclusion or affinity chromatographies will be performed.

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RUPRECHT-KARLS-UNIVERSITAET HEIDELBERG
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69120 HEIDELBERG
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