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非洲爪蟾卵母细胞中一个翻译控制的信使核糖核酸的破坏会延迟孕酮诱导的成熟。

Destruction of a translationally controlled mRNA in Xenopus oocytes delays progesterone-induced maturation.

作者信息

Smith R C, Dworkin M B, Dworkin-Rastl E

机构信息

Ernst-Boehringer-Institut, Vienna, Austria.

出版信息

Genes Dev. 1988 Oct;2(10):1296-306. doi: 10.1101/gad.2.10.1296.

Abstract

The maternal mRNA D7 is a moderately abundant transcript in Xenopus laevis whose expression is highest in, and perhaps restricted to, oogenesis and early embryogenesis. The nucleotide sequence of cloned D7 cDNA was determined and shown to have the capacity to code for a 31-kD protein. This amino acid sequence was searched against a protein data base, and no homologous proteins were found. Antibodies directed against D7 recognize in Xenopus embryos a soluble, cytoplasmic protein with an apparent molecular weight on SDS gels of 36,000. The D7 protein is absent from oocytes and first begins to accumulate during oocyte maturation. Its levels are highest during the first day of embryonic development and then decrease; D7 protein was not detected in adult tissues. D7 mRNA was selectively destroyed by injection into oocytes of antisense oligodeoxynucleotides. Analysis of injected oocytes by Northern and Western blotting showed site-specific cleavage and subsequent degradation of the D7 mRNA and the failure of the D7 protein to accumulate during progesterone-induced maturation. The loss of D7 protein affects the maturation process itself, significantly delaying the time course of germinal vesicle breakdown. Thus, D7 is a newly described protein involved in oocyte maturation.

摘要

母体mRNA D7是非洲爪蟾中一种中等丰度的转录本,其表达在卵子发生和早期胚胎发生过程中最高,可能也仅限于此阶段。已确定克隆的D7 cDNA的核苷酸序列,显示其有能力编码一种31-kD的蛋白质。将该氨基酸序列与蛋白质数据库进行比对,未发现同源蛋白质。针对D7的抗体在非洲爪蟾胚胎中识别出一种可溶性细胞质蛋白,在SDS凝胶上的表观分子量为36,000。卵母细胞中不存在D7蛋白,它在卵母细胞成熟过程中开始积累。其水平在胚胎发育的第一天最高,然后下降;在成体组织中未检测到D7蛋白。通过向卵母细胞注射反义寡脱氧核苷酸选择性地破坏了D7 mRNA。通过Northern和Western印迹分析注射后的卵母细胞,显示D7 mRNA发生位点特异性切割并随后降解,且在孕酮诱导的成熟过程中D7蛋白未能积累。D7蛋白的缺失影响成熟过程本身,显著延迟生发泡破裂的时间进程。因此,D7是一种新描述的参与卵母细胞成熟的蛋白质。

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