Walker J, Minshall N, Hake L, Richter J, Standart N
Department of Biochemistry, University of Cambridge, United Kingdom.
RNA. 1999 Jan;5(1):14-26. doi: 10.1017/s1355838299981219.
During early development gene expression is controlled principally at the translational level. Oocytes of the surf clam Spisula solidissima contain large stockpiles of maternal mRNAs that are translationally dormant or masked until meiotic maturation. Activation of the oocyte by fertilization leads to translational activation of the abundant cyclin and ribonucleotide reductase mRNAs at a time when they undergo cytoplasmic polyadenylation. In vitro unmasking assays have defined U-rich regions located approximately centrally in the 3' UTRs of these mRNAs as translational masking elements. A clam oocyte protein of 82 kDa, p82, which selectively binds the masking elements, has been proposed to act as a translational repressor. Importantly, mRNA-specific unmasking in vitro occurs in the absence of poly(A) extension. Here we show that clam p82 is related to Xenopus CPEB, an RNA-binding protein that interacts with the U-rich cytoplasmic polyadenylation elements (CPEs) of maternal mRNAs and promotes their polyadenylation. Cloned clam p82/CPEB shows extensive homology to Xenopus CPEB and related polypeptides from mouse, goldfish, Drosophila and Caenorhabditis elegans, particularly in their RNA-binding C-terminal halves. Two short N-terminal islands of sequence, of unknown function, are common to vertebrate CPEBs and clam p82. p82 undergoes rapid phosphorylation either directly or indirectly by cdc2 kinase after fertilization in meiotically maturing clam oocytes, prior to its degradation during the first cell cleavage. Phosphorylation precedes and, according to inhibitor studies, may be required for translational activation of maternal mRNA. These data suggest that clam p82 may be a functional homolog of Xenopus CPEB.
在早期发育过程中,基因表达主要在翻译水平受到调控。硬壳蛤(Spisula solidissima)的卵母细胞含有大量母源mRNA储备,这些mRNA在减数分裂成熟之前处于翻译休眠或被掩盖状态。受精激活卵母细胞会导致大量细胞周期蛋白和核糖核苷酸还原酶mRNA在经历细胞质多聚腺苷酸化时发生翻译激活。体外去掩盖试验已确定这些mRNA的3'UTR大约位于中央的富含U的区域为翻译掩盖元件。一种82 kDa的蛤卵母细胞蛋白p82,它选择性地结合掩盖元件,被认为是一种翻译抑制因子。重要的是,体外mRNA特异性去掩盖在没有多聚腺苷酸化延伸的情况下发生。在这里我们表明,蛤p82与非洲爪蟾CPEB相关,CPEB是一种RNA结合蛋白,它与母源mRNA的富含U的细胞质多聚腺苷酸化元件(CPE)相互作用并促进其多聚腺苷酸化。克隆的蛤p82/CPEB与非洲爪蟾CPEB以及来自小鼠、金鱼、果蝇和秀丽隐杆线虫的相关多肽具有广泛的同源性,特别是在它们的RNA结合C端部分。脊椎动物CPEB和蛤p82有两个功能未知的短N端序列岛是共同的。在减数分裂成熟的蛤卵母细胞受精后,p82在第一次细胞分裂期间降解之前,会被cdc2激酶直接或间接快速磷酸化。磷酸化先于母源mRNA翻译激活发生,并且根据抑制剂研究,可能是其翻译激活所必需的。这些数据表明,蛤p82可能是非洲爪蟾CPEB的功能同源物。