A role for cyclin A1 in the activation of MPF and G2-M transition during meiosis of male germ cells in mice.
Cell-cycle transition at G2-M is controlled by MPF (M-phase-promoting factor), a complex consisting of the Cdc2 kinase and a B-type cyclin. We have shown that in mice, targeted disruption of an A-type cyclin gene, cyclin A1, results in a block of spermatogenesis prior to the entry into metaphase I. The meiotic arrest is accompanied by a defect in Cdc2 kinase activation at the G2--M transition, raising the possibility that a cyclin A1-dependent process dictates the activation of MPF. Here we show that like Cdc2, the expression of B-type cyclins is retained in cyclin A1-deficient spermatocytes, while their associated kinases are kept at inactive states. Treatment of arrested germ cells with the protein phosphatase type-1 and -2A inhibitor okadaic acid restores the MPF activity and induces entry into M phase and the formation of normally condensed chromosome bivalents, concomitant with hyperphosphorylation of Cdc25 proteins. Conversely, inhibition of tyrosine phosphatases, including Cdc25s, by vanadate suppresses the okadaic acid-induced metaphase induction. The highest levels of Cdc25A and Cdc25C expression and their subcellular localization during meiotic prophase coincide with that of cyclin A1, and when overexpressed in HeLa cells, cyclin A1 coimmunoprecipitates with Cdc25A. Furthermore, the protein kinase complexes consisting of cyclin A1 and either Cdc2 or Cdk2 phosphorylate both Cdc25A and Cdc25C in vitro. These results suggest that in normal meiotic male germ cells, cyclin A1 participates in the regulation of other protein kinases or phosphatases critical for the G2-M transition. In particular, it may be directly involved in the initial amplification of MPF through the activating phosphorylation on Cdc25 phosphatases.
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The Integrated Program in Cellular, Molecular and Biophysical Studies, Columbia University College of Physicians and Surgeons, New York, NY 10032, USA. djw3@columbia.edu
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Wolgemuth
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D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D019925:Cyclin A / Q000502:physiology*; D056750:Cyclin A1; D017931:DNA Primers; D016195:G2 Phase; D008297:Male; D016200:Maturation-Promoting Factor / Q000378:metabolism*; D008540:Meiosis; D000090204:Mesothelin; D051379:Mice; D008810:Mice, Inbred C57BL; D008938:Mitosis; D019319:Okadaic Acid / Q000494:pharmacology; D010766:Phosphorylation; D013090:Spermatocytes / Q000166:cytology*; D013737:Testis / Q000166:cytology / Q000187:drug effects; D014443:Tyrosine / Q000378:metabolism
C503747:Ccna1 protein, mouse; D019925:Cyclin A; D056750:Cyclin A1; D017931:DNA Primers; C000715528:Msln protein, mouse; D019319:Okadaic Acid; D014443:Tyrosine; D016200:Maturation-Promoting Factor; D000090204:Mesothelin