A novel p34(cdc2)-binding and activating protein that is necessary and sufficient to trigger G(2)/M progression in Xenopus oocytes.
The activation of maturation-promoting factor (MPF) is required for G(2)/M progression in eukaryotic cells. Xenopus oocytes are arrested in G(2) and are induced to enter M phase of meiosis by progesterone stimulation. This process is known as meiotic maturation and requires the translation of specific maternal mRNAs stored in the oocytes. We have used an expression cloning strategy to functionally identify proteins involved in G(2)/M progression in Xenopus oocytes. Here we report the cloning of two novel cDNAs that when expressed in oocytes induce meiotic maturation efficiently. The two cDNAs encode proteins of 33 kD that are 88% identical and have no significant homologies to other sequences in databases. These proteins, which we refer to as p33(ringo) (rapid inducer of G(2)/M progression in oocytes), induce very rapid MPF activation in cycloheximide-treated oocytes. Conversely, ablation of endogenous p33(ringo) mRNAs using antisense oligonucleotides inhibits progesterone-induced maturation, suggesting that synthesis of p33(ringo) is required for this process. We also show that p33(ringo) binds to and activates the kinase activity of p34(cdc2) but does not associate with p34(cdc2)/cyclin B complexes. Our results identify a novel p34(cdc2) binding and activating protein that regulates the G(2)/M transition during oocyte maturation.
Ferby
I
I
European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
Blazquez
M
M
Palmer
A
A
Eritja
R
R
Nebreda
A R
AR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D019926:Cyclin B / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D018076:DNA, Complementary; D004789:Enzyme Activation; D016195:G2 Phase; D016200:Maturation-Promoting Factor / Q000378:metabolism; D008938:Mitosis; D008969:Molecular Sequence Data; D009865:Oocytes / Q000166:cytology / Q000187:drug effects; D011374:Progesterone / Q000494:pharmacology; D011817:Rabbits; D014981:Xenopus; D029867:Xenopus Proteins
D018797:Cell Cycle Proteins; D019926:Cyclin B; D018076:DNA, Complementary; D029867:Xenopus Proteins; C119055:ls27 protein, Xenopus; D011374:Progesterone; D003513:Cycloheximide; D016203:CDC2 Protein Kinase; D016200:Maturation-Promoting Factor