Lanclos K D, Hamilton T H
Proc Natl Acad Sci U S A. 1975 Oct;72(10):3934-8. doi: 10.1073/pnas.72.10.3934.
Induction of the synthesis of the vitellogenic proteins, lipovitellin and phosvitin, in the liver of the male African clawed toad (Xenopus laevis) was investigated as a function of time after treatment with estradiol-17beta [1,3,5(10)-estratriene-3,17beta-diol]. The appearance of mRNAs encoded for lipovitellin and phosvitin in the cytoplasmic fraction of the liver was assayed by microinjections of hepatic mRNA preparation [either polyribosomes or poly(A)-rich RNA] into oocytes obtained from mature female toads. Oocytes were then incubated in the presence of radioactive amino acid(s) at 19 degrees for periods of time varying from 4 to 18 hr after microinjection. The results show that at 2 hr after hormone treatment more mRNA was present in the cytoplasm, and that from 2 to 72 hr after treatment the level of induced mRNA increased almost linearly to 110% above the control values. Experiments employing specific lipovitellin antiserum indicated no radioactive lipovitellin among the proteins synthesized in oocytes microinjected with hepatic mRNAs isolated from 3 to 9 hr after hormone treatment. However, a marked synthesis of immunoprecipitable, radioactive lipovitellin and an enhanced incorporation of [3H]serine occurred in the oocytes microinjected with hepatic mRNA preparations obtained from toads treated with hormone for 12 or more hr. The identities of the proteins encoded by the mRNAs induced early in estrogen action (2-9 hr) in the male amphibian liver are unknown. It is surmised that some of these proteins may function in the regulation of the subsequent synthesis of the vitellogenic proteins.
研究了用17β-雌二醇[1,3,5(10)-雌三烯-3,17β-二醇]处理后,雄性非洲爪蟾(非洲爪蟾)肝脏中卵黄生成蛋白、脂磷蛋白和卵黄高磷蛋白合成的诱导情况与时间的关系。通过将肝脏mRNA制剂(多核糖体或富含多聚腺苷酸的RNA)显微注射到从成熟雌性蟾蜍获得的卵母细胞中,来检测肝脏细胞质部分中脂磷蛋白和卵黄高磷蛋白编码的mRNA的出现情况。然后将卵母细胞在放射性氨基酸存在下于19℃孵育,显微注射后孵育4至18小时不等。结果表明,激素处理后2小时,细胞质中存在更多的mRNA,并且在处理后2至72小时,诱导的mRNA水平几乎呈线性增加,比对照值高110%。使用特异性脂磷蛋白抗血清的实验表明,在用激素处理3至9小时后分离的肝脏mRNA显微注射的卵母细胞中合成的蛋白质中没有放射性脂磷蛋白。然而,在用激素处理12小时或更长时间的蟾蜍获得的肝脏mRNA制剂显微注射的卵母细胞中,出现了明显的可免疫沉淀的放射性脂磷蛋白合成以及[3H]丝氨酸掺入增加的情况。雄性两栖动物肝脏中雌激素作用早期(2至9小时)诱导的mRNA编码的蛋白质的身份尚不清楚。据推测,其中一些蛋白质可能在随后的卵黄生成蛋白合成的调节中发挥作用。