Evidence that Mos protein may not act directly on cyclin.
Using affinity-purified antiserum we have examined cyclin B2 levels in Xenopus oocytes at various stages of oogenesis. We found that cyclin B2 is detected from stage 2 to stage 6 as two bands, one of which is phosphorylated, and that cyclin B2 mass increases about 28-fold between stage 2 and stage 6. To examine the effect of Mos protein on cyclin phosphorylation, we microinjected synthetic Xenopus c-mos (c-mosxe) RNA into stages 4, 5, and 6 Xenopus oocytes. In stage 6 oocytes, maturation was induced by c-mosxe RNA, and, as is the case with progesterone treatment, all cyclin B2 was shifted to the phosphorylated form. However, c-mosxe RNA injected into stage 4 or 5 oocytes did not induce maturation or cause a shift in the relative proportion of the two cyclin B2 bands. These data suggest that Mos does not act directly to phosphorylate cyclin B2, causing the band shift during maturation. Cyclin B2 synthesis increases about 2-fold during maturation, in concert with total protein synthesis. Data from experiments on cyclin B2 stability indicate that the half-life of cyclin B2 is about 85 hr in stage 6. This suggests that if Mos protein has a direct effect on cyclin stability, it does so only at a later stage in oocyte maturation, but not at the onset.
Xu
W
W
Department of Developmental and Cell Biology, University of California, Irvine 92717.
Ladner
K J
KJ
Smith
L D
LD
D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D016213:Cyclins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D016258:Genes, mos; D007700:Kinetics; D008845:Microinjections; D009865:Oocytes / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D017231:Proto-Oncogene Proteins c-mos; D012313:RNA / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014982:Xenopus laevis
D016213:Cyclins; D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins; D012313:RNA; D011505:Protein-Tyrosine Kinases; D017231:Proto-Oncogene Proteins c-mos