A member of the Ste20/PAK family of protein kinases is involved in both arrest of Xenopus oocytes at G2/prophase of the first meiotic cell cycle and in prevention of apoptosis.
We have identified new members (X-PAKs) of the Ste20/PAK family of protein kinases in Xenopus, and investigated their role in the process that maintains oocytes arrested in the cell cycle. Microinjection of a catalytically inactive mutant of X-PAK1 with a K/R substitution in the ATP binding site, also deleted of its Nter-half that contains the conserved domains responsible for binding of both Cdc42/Rac GTPases and SH3-containing proteins, greatly facilitates oocyte release from G2/prophase arrest by progesterone and insulin. Addition of the same X-PAK1 mutant to cell cycle extracts from unfertilized eggs induced apoptosis, as shown by activation of caspases and cytological changes in in vitro-assembled nuclei. This was suppressed by adding Bcl-2 or the DEVD peptide inhibitor of caspases, and rescued by competing the dominant-negative mutant with its constitutively active X-PAK1 counterpart. Such results indicate that X-PAK1 (or another member of the Xenopus Ste20/PAK family of protein kinases) is involved in arrest of oocytes at G2/prophase and prevention of apoptosis; thus death by apoptosis and release of healthy oocytes from cell cycle arrest may be linked. That cell cycle arrest protects oocytes from apoptosis is consistent with the finding that extracts from metaphase II-arrested oocytes are less sensitive to apoptotic signals than those from activated eggs.
Faure
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Centre de Recherches de Biochimie Macromoléculaire, CNRS ERS 155, 1919 route de Mende, 34293 Montpellier Cedex 5, France.
Vigneron
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Dorée
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Morin
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N
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001483:Base Sequence; D001665:Binding Sites; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D017931:DNA Primers; D005260:Female; D016195:G2 Phase; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins; D007700:Kinetics; D020930:MAP Kinase Kinase Kinases; D008540:Meiosis / Q000187:drug effects / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009842:Oligopeptides / Q000494:pharmacology; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011374:Progesterone / Q000494:pharmacology; D011418:Prophase; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014981:Xenopus; D054462:p21-Activated Kinases
D017931:DNA Primers; D007328:Insulin; D047908:Intracellular Signaling Peptides and Proteins; D009842:Oligopeptides; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D011374:Progesterone; D000255:Adenosine Triphosphate; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases; D020930:MAP Kinase Kinase Kinases; C078754:STE20 protein, S cerevisiae