Taylor M A, Johnson A D, Smith L D
Proc Natl Acad Sci U S A. 1985 Oct;82(19):6586-9. doi: 10.1073/pnas.82.19.6586.
Previous studies have shown that exogenous mRNAs injected into full-grown (stage 6) Xenopus oocytes are translated only at the expense of endogenous messages; translational capacity is limited. In this report, we demonstrate that injection of globin mRNA into small, stage 4 oocytes results in an increase in total protein synthesis without a concomitant decrease in the translation of endogenous mRNAs. The absence of competition with endogenous messages in stage 4 oocytes, injected with globin mRNA, compared with stage 6 oocytes, was not due to differential turnover of the injected mRNA. Hybridization of RNA from mRNA-injected oocytes at both stages revealed that similar amounts of globin mRNA were present. These results are interpreted to mean that protein synthesis in growing oocytes is limited by the availability of mRNA and not components of the translational machinery. This conclusion, however, does not apply to all mRNA classes. The capacity of stage 4 oocytes to translate a mRNA (zein) on membrane-bound polysomes is as restricted as reported previously for stage 6 oocytes. This result suggests that putative membrane binding sites or rough endoplasmic reticulum content are limited in oocytes at both stages. The possible role of association of injected mRNA with endogenous proteins that prevent translation is discussed.
先前的研究表明,注射到完全成熟(第6阶段)非洲爪蟾卵母细胞中的外源mRNA仅以内源信息为代价进行翻译;翻译能力是有限的。在本报告中,我们证明将珠蛋白mRNA注射到小型的第4阶段卵母细胞中会导致总蛋白合成增加,而内源mRNA的翻译不会随之减少。与第6阶段卵母细胞相比,注射了珠蛋白mRNA的第4阶段卵母细胞中不存在与内源信息的竞争,这并非由于注射的mRNA的不同周转率所致。对两个阶段注射了mRNA的卵母细胞的RNA进行杂交分析表明,存在相似量的珠蛋白mRNA。这些结果被解释为意味着正在生长的卵母细胞中的蛋白质合成受mRNA可用性的限制,而非翻译机制的组成部分。然而,这一结论并不适用于所有mRNA类别。第4阶段卵母细胞在膜结合多聚核糖体上翻译一种mRNA(玉米醇溶蛋白)的能力与先前报道的第6阶段卵母细胞一样受到限制。这一结果表明,两个阶段的卵母细胞中假定的膜结合位点或粗面内质网含量都是有限的。本文还讨论了注射的mRNA与阻止翻译的内源蛋白结合的可能作用。