Tyrosine phosphorylation of p34cdc2 and p42 during meiotic maturation of Xenopus oocyte. Antagonistic action of okadaic acid and 6-DMAP.
The tyrosine phosphorylation/dephosphorylation of p34cdc2 was estimated by immunoblotting with antiphosphotyrosine antibody during meiotic maturation of Xenopus oocytes. At the time of germinal vesicle breakdown (GVBD), p34cdc2 is tyrosine dephosphorylated whereas a p42 protein, which might correspond to a MAP2 kinase, becomes tyrosine phosphorylated. No modification in the level of tyrosine phosphorylation of either proteins was noticed during the whole maturation process from GVBD until metaphase II. When added to prophase oocytes, 6-DMAP (6-dimethyl-aminopurine) blocks GVBC, M-phase-promoting factor (MPF) activation and H1-histone, kinase activation induced by either progesterone, MPF transfer or okadaic acid microinjection. In each case, the tyrosine dephosphorylation reaction of p34cdc2 is inhibited. In meiosis I oocytes (just after the initiation of GVBD), 6-DMAP provokes the rephosphorylation of p34cdc2 on tyrosine residue(s), inactivation of MPF and H1-histone kinase and re-entry of the cell into an interphase-like state. These processes are reversible by simply removing the agent. In contrast to the observations in prophase oocytes, okadaic acid is able to reverse the inhibitory effect of 6-DMAP in meiosis I oocytes on MPF and H1-histone kinase activities and to initiate dephosphorylation of p34cdc2 on tyrosyl residue(s) even in the presence of 6-DMAP. Altogether, our results show that 6-DMAP and okadaic acid antagonistically control in vivo the level of tyrosine phosphorylation of p34cdc2.
Jessus
C
C
Laboratoire de Physiologie de la Reproduction, UA-CNRS/INRA 555, Université P. et M. Curie, Paris, France.
Rime
H
H
Haccard
O
O
Van Lint
J
J
Goris
J
J
Merlevede
W
W
Ozon
R
R
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D004988:Ethers, Cyclic / Q000494:pharmacology; D005260:Female; D015151:Immunoblotting; D007476:Ionophores / Q000494:pharmacology; D016200:Maturation-Promoting Factor / Q000502:physiology*; D008540:Meiosis / Q000502:physiology*; D019319:Okadaic Acid; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011506:Proteins / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*; D014982:Xenopus laevis
D004988:Ethers, Cyclic; D007476:Ionophores; D011506:Proteins; D019319:Okadaic Acid; D014443:Tyrosine; C001020:N(6),N(6)-dimethyladenine; D016203:CDC2 Protein Kinase; D016200:Maturation-Promoting Factor; D000225:Adenine