Varnum S M, Hurney C A, Wormington W M
Department of Biology, Brandeis University, Waltham, Massachusetts 02254.
Dev Biol. 1992 Oct;153(2):283-90. doi: 10.1016/0012-1606(92)90113-u.
During the meiotic maturation of Xenopus oocytes, maternal mRNAs that lack a cytoplasmic polyadenylation element are deadenylated and translationally inactivated. In this report, we have characterized the regulation of poly(A) removal during maturation. Deadenylation in vivo is detected only after germinal vesicle breakdown and does not require de novo protein synthesis. Enucleated oocytes do not deadenylate either endogenous or microinjected RNAs upon maturation, indicating that a nuclear component is required for poly(A) removal. Whole cell extracts prepared from both immature and mature oocytes deadenylate exogenous RNA substrates in vitro. Deadenylation activity is not detected in isolated nuclear or cytoplasmic extracts obtained from immature oocytes, but is reconstituted when these fractions are combined in vitro. These results indicate that the factors required for deadenylation activity are present in immature oocytes, but that poly(A) removal is prevented by the sequestration of one or more of these components within the nucleus. Maturation-specific deadenylation of maternal mRNAs occurs upon the release of nuclear factors into the cytoplasm at germinal vesicle breakdown.
在非洲爪蟾卵母细胞的减数分裂成熟过程中,缺乏细胞质聚腺苷酸化元件的母源mRNA会发生去腺苷酸化并在翻译水平上失活。在本报告中,我们对成熟过程中聚腺苷酸去除的调控进行了表征。体内去腺苷酸化仅在生发泡破裂后才被检测到,且不需要从头合成蛋白质。去核卵母细胞在成熟时不会使内源性或显微注射的RNA发生去腺苷酸化,这表明聚腺苷酸的去除需要一种核成分。从未成熟和成熟卵母细胞制备的全细胞提取物在体外会使外源RNA底物发生去腺苷酸化。从未成熟卵母细胞获得的分离核提取物或细胞质提取物中未检测到去腺苷酸化活性,但当这些组分在体外混合时,去腺苷酸化活性得以重建。这些结果表明,去腺苷酸化活性所需的因子存在于未成熟卵母细胞中,但聚腺苷酸的去除因这些组分中的一种或多种被隔离在细胞核内而受到抑制。母源mRNA的成熟特异性去腺苷酸化发生在生发泡破裂时核因子释放到细胞质中之后。