Cyclin B in fish oocytes: Its cDNA and amino acid sequences, appearance during maturation, and induction of p34^cdc2 activation

Under the influence of maturation‐inducing hormone (MIH) secreted from follicle cells, oocyte maturation is finally triggered by maturation‐promoting factor (MPF), which consists of a homolog of the cdc2⁺ gene product of fission yeast (p34^cdc2) and cyclin B. Two species of cyclin B clones were isolated from a cDNA library constructed from mature goldfish oocytes. Sequence comparisons revealed that these two clones are highly homologous (95%) and were found to be similar to Xenopus cyclin B1. Using monocional antibodies against Escherichia coli produced goldfish cyclin B and the PSTAIR sequence of p34^cdc2, we examined the levels of cyclin B and p34^cdc2 proteins during goldfish oocyte maturation induced in vitro by 17α,20β‐dihydroxy‐4‐pregnen‐3‐one (17α,20β‐DP), a natural MIH in fish. Protein p34^cdc2 was found in immature oocyte extracts and did not remarkably change during oocyte maturation. Cyclin B was not detected in immature oocyte extracts and appeared when oocytes underwent germinal vesicle breakdown. Cyclin B that appeared during oocyte maturation was labelled with [³⁵S]methionine, indicating its de novo synthesis. Introduction of E. coli‐produced cyclin B into immature oocyte extracts induced p34^cdc2 (MPF) activation. Although the possibility that immature goldfish oocytes contain an insoluble cyclin B is not completely excluded, these results strongly suggest that 17α, 20β‐DP induced oocytes to synthesize cyclin B, which in turn activates preexisting p34^cdc2, forming active MPF. © 1992 Wiley‐Liss, Inc.