Sierra J M, Meier D, Ochoa S
Proc Natl Acad Sci U S A. 1974 Jul;71(7):2693-7. doi: 10.1073/pnas.71.7.2693.
Cell-free preparations from encapsulated Artemia salina embryos readily translate poly(U), but there is no endogenous protein synthesis or translation of added natural mRNA until development resumes upon incubation in saline. High-salt-washed 80S ribosomes and highspeed supernatant (cytosol) were prepared from undeveloped eggs and from eggs allowed to develop to a stage prior to hatching. Endogenous protein synthesis occurs only with ribosomes from developed embryos, whether with undeveloped or developed cytosol. This is mainly elongation of preformed polypeptide chains, for it is largely resistant to edeine, an inhibitor of chain initiation. Edeinesensitive translation of added natural mRNA occurs with both developed and undeveloped ribosomes but requires developed cytosol. Translation of brome mosaic virus RNA is not much, if at all, further stimulated by high-saltwash of developed ribosomes, but that of globin mRNA is markedly enhanced. Levels of the chain initiation factor EIF-1 are the same before and after development. These results are consistent with the view that resumption of developemtn is triggered by transcription, with ensuing translation of the resulting messengers to yield, among other proteins, mRNA-recognizing initiation factors of the IF-3 type which are partly free in the cytosol and partly ribosome-bound. The data also suggest that different factors may be involved in the translation of brome mosaic virus RNA and globin mRNA by this system.
来自包囊卤虫胚胎的无细胞制剂能够轻易地翻译聚尿苷酸,但在盐溶液中孵育直至发育恢复之前,不存在内源性蛋白质合成或对添加的天然信使核糖核酸(mRNA)的翻译。从未发育的卵以及发育至孵化前阶段的卵中制备了经高盐洗涤的80S核糖体和高速上清液(胞质溶胶)。内源性蛋白质合成仅发生在来自已发育胚胎的核糖体中,无论使用未发育的还是已发育的胞质溶胶。这主要是预先形成的多肽链的延伸,因为它在很大程度上对放线菌素(一种链起始抑制剂)具有抗性。添加的天然mRNA的放线菌素敏感翻译在已发育和未发育的核糖体中均会发生,但需要已发育的胞质溶胶。已发育核糖体的高盐洗涤对雀麦花叶病毒RNA的翻译即使有刺激作用也不大,但对珠蛋白mRNA的翻译有显著增强。发育前后链起始因子EIF-1的水平相同。这些结果与以下观点一致,即发育的恢复是由转录触发的,随后产生的信使核糖核酸被翻译以产生除其他蛋白质外的IF-3型信使核糖核酸识别起始因子,这些因子部分游离于胞质溶胶中,部分与核糖体结合。数据还表明,该系统翻译雀麦花叶病毒RNA和珠蛋白mRNA可能涉及不同的因子。