role for non-proteolytic control of m-phase promoting factor activity at m-phase exit角色non-proteolytic活动在有丝分裂期出口控制有丝分裂期的促进因素.pdfVIP

Role for Non-Proteolytic Control of M-phase Promoting Factor Activity at M-phase Exit 1,3 1,2 1,2 4 1,2 Vincenzo D’Angiolella , Luca Palazzo , Concetta Santarpia , Vincenzo Costanzo , Domenico Grieco * 1 Faculty of Biotechnological Sciences and Dipartimento di Biologia e Patologia Cellulare e Molecolare ‘‘L. Califano,’’ University of Napoli Federico II, Italy, 2 CEINGE Biotecnologie Avanzate, Napoli, Italy, 3 Department of Pathology, New York University, New York, United States of America, 4 Clare Hall Laboratories, London Research Institute, London, United Kingdom M-phase Promoting Factor (MPF; the cyclin B-cdk 1 complex) is activated at M-phase onset by removal of inhibitory phosphorylation of cdk1 at thr-14 and tyr-15. At M-phase exit, MPF is destroyed by ubiquitin-dependent cyclin proteolysis. Thus, control of MPF activity via inhibitory phosphorylation is believed to be particularly crucial in regulating transition into, rather than out of, M-phase. Using the in vitro cell cycle system derived form Xenopus eggs, here we show, however, that inhibitory phosphorylation of cdk1 contributes to control MPF activity during M-phase exit. By sampling extracts at very short intervals during both meiotic and mitotic exit, we found that cyclin B1-associated cdk1 underwent transient inhibitory phosphorylation at tyr-15 and that cyclin B1-cdk1 activity fell more rapidly than the cyclin B1 content. Inhibitory phosphorylation of MPF correlated with phosphorylation changes of cdc25C, the MPF phosphatase, and physical interaction of cdk1 with wee1, the MPF kinase, during M-phase exit. MPF down-regulation required Ca++/calmodulin-dependent kinase II (CaMKII) and cAMP-dependent protein kinase