Maturation of Xenopus laevis oocyte by progesterone requires poly(A) tail elongation of mRNA.
Meiotic maturation of Xenopus laevis oocytes by progesterone requires translation of stored maternal mRNAs. We investigated the role of poly(A) tail elongation of mRNAs during this process using cordycepin, which inhibits poly(A) tail elongation of mRNAs. When oocytes were treated with the buffer containing 10 mM cordycepin for 12 h, concentration of 3'-dATP in cytosol of oocytes increased to 0.7 mM, while that of ATP remained constant at around 1.2 mM. Incorporation of [32P]AMP into poly(A) mRNA was inhibited almost completely by this treatment. Progesterone-induced germinal vesicle breakdown (GVBD) was also abolished. Dose dependence of inhibition of progesterone-induced GVBD on cordycepin was similar to that of [32P]AMP incorporation into poly(A) mRNA. However, maturation-promoting factor-induced GVBD was unaffected by treatment of oocytes with cordycepin. Furthermore, the inhibition of GVBD by cordycepin was rescued by removal of cordycepin even in the presence of actinomycin D. Therefore, we concluded that poly(A) tail elongation of mRNA is required for induction of meiotic maturation of X. laevis oocytes. In addition, progesterone induced a 2.7-fold activation of [32P]AMP incorporation into the poly(A) tail of mRNA after a lag period of 3 h whereas GVBD was induced after 6-8 h from the progesterone treatment. Syntheses of most of the proteins were unaffected by treatment of oocytes with progesterone or cordycepin. However, syntheses of several proteins were increased or decreased by progesterone and cordycepin treatment.
Kuge
H
H
Department of Biology, Faculty of Science, Osaka University, Japan.
Inoue
A
A
D000249:Adenosine Monophosphate / Q000378:metabolism*; D000818:Animals; D003513:Cycloheximide / Q000494:pharmacology; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D003839:Deoxyadenosines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D066298:In Vitro Techniques; D016200:Maturation-Promoting Factor / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010761:Phosphorus Radioisotopes; D011061:Poly A / Q000378:metabolism*; D011374:Progesterone / Q000494:pharmacology*; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism*; D014982:Xenopus laevis
D003838:Deoxyadenine Nucleotides; D003839:Deoxyadenosines; D010761:Phosphorus Radioisotopes; D011506:Proteins; D012333:RNA, Messenger; D011061:Poly A; D000249:Adenosine Monophosphate; D011374:Progesterone; D003513:Cycloheximide; D008715:Methionine; D016200:Maturation-Promoting Factor; C058120:cordycepin; C026600:2'-deoxyadenosine triphosphate