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卤虫早期胚胎发育过程中的信使核糖核酸:翻译能力的改变与序列复杂性

Messenger RNA during early embryogenesis in Artemia salina: altered translatability and sequence complexity.

作者信息

James T C, Tata J R

出版信息

Differentiation. 1980 Feb;16(1):9-21. doi: 10.1111/j.1432-0436.1980.tb01053.x.

Abstract

RNA from developing embryos of Artemia salina (5, 10, and 20 h after re-initiation of development) was translated 3-10 times more efficiently in a rabbit reticulocyte lysate cell-free protein synthesizing system than RNA from dormant gastrulae. The latter did not appear to contain any significant amount of translation inhibitor activity. Ninety percent of the translatable activity in dormant gastrulae was recovered as poly(A)--RNA, whereas 80% of that in post-gastrular developing embryos was present as poly(A)+-RNA. The size of most polypeptides coded for by dormant gastrular RNA was less than 130,000 daltons whereas the size of those coded for by developing embryonic RNA was up to 200,000 daltons, which correlated with a corresponding shift to poly A-containing RNA of higher molecular weight. Two major polypeptides of about 37,000 daltons coded for by dormant gastrular RNA disappeared at 20 h after resumption of development. Hybridization of complementary DNA (cDNA) to a 1000-fold excess of the homologous poly(A)+-RNA revealed the presence of three complexity classes of mRNA. Forty-five percent, 30%, and 25% of RNA in dormant gastrulae were present as high, middle, and low abundance classes comprising about 10, 80, and 9700 species, respectively whereas in the nauplii there were 10, 150, and 7900 species of high, middle, and low abundancy sequences, respectively. Heterologous hybridizations using cDNA complementary to highly abundant messenger population of nauplii (isolated by chromatography on hydroxyapatite) to poly(A)+-RNA from dormant cysts showed considerably divergence in this class of messengers from the two developmental stages. Re-initiation of development of dormant Artemia gastrulae is thus characterized by a "re-programming" seen as a simultaneous and rapid increase in the polyadenylation and translatability of poly(A)+-RNA accompanied by a qualitative change in its sequence complexity.

摘要

卤虫发育胚胎(发育重新启动后5、10和20小时)的RNA在兔网织红细胞裂解物无细胞蛋白质合成系统中的翻译效率比休眠原肠胚的RNA高3至10倍。后者似乎不含有任何显著量的翻译抑制活性。休眠原肠胚中90%的可翻译活性以聚腺苷酸(poly(A))-RNA的形式回收,而原肠胚后发育胚胎中80%的可翻译活性以聚腺苷酸(poly(A)+)-RNA的形式存在。休眠原肠胚RNA编码的大多数多肽大小小于130,000道尔顿,而发育胚胎RNA编码的多肽大小可达200,000道尔顿,这与向更高分子量的含聚腺苷酸RNA的相应转变相关。休眠原肠胚RNA编码的两种约37,000道尔顿的主要多肽在发育恢复后20小时消失。互补DNA(cDNA)与1000倍过量的同源聚腺苷酸(poly(A)+)-RNA杂交揭示了三类复杂程度不同的mRNA的存在。休眠原肠胚中45%、30%和25%的RNA分别以高、中、低丰度类别存在,分别包含约10、80和9700种,而在无节幼体中,高、中、低丰度序列分别有10、150和7900种。使用与无节幼体高度丰富信使群体互补的cDNA(通过羟基磷灰石层析分离)与休眠囊泡的聚腺苷酸(poly(A)+)-RNA进行异源杂交,显示这两个发育阶段的这类信使存在相当大的差异。因此,休眠卤虫原肠胚发育的重新启动的特征是一种“重新编程”,表现为聚腺苷酸(poly(A)+)-RNA的聚腺苷酸化和可翻译性同时快速增加,并伴随着其序列复杂性的质的变化。

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