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非洲爪蟾早期发育过程中Na(+)-K(+)-ATP酶β1亚基的多聚腺苷酸化

Polyadenylation of Na(+)-K(+)-ATPase beta 1-subunit during early development of Xenopus laevis.

作者信息

Burgener-Kairuz P, Corthesy-Theulaz I, Merillat A M, Good P, Geering K, Rossier B C

机构信息

Institut de Pharmacologie et de Toxicologie, Université de Lausanne, Switzerland.

出版信息

Am J Physiol. 1994 Jan;266(1 Pt 1):C157-64. doi: 10.1152/ajpcell.1994.266.1.C157.

Abstract

In fully grown Xenopus oocytes, the synthesis of beta-subunits is limiting for the formation of functional Na(+)-K(+)-adenosinetriphosphatase alpha/beta-complexes (Geering, K. FEBS Lett. 285: 189-193, 1991). In the present study, we show that during oocyte growth (from stage I to stage VI) alpha 1-, but not beta 1- or beta 3-isoform, mRNAs accumulate. In addition, beta-mRNAs are apparently sequestered in an untranslated pool in fully grown oocytes (stage VI). From fertilization to morulation, the total pools of alpha 1-, beta 1-, or beta 3-mRNAs vary little. Whereas polyadenylated [poly(A)+] alpha 1- and beta 3-isoform mRNAs did not change significantly, poly(A)+ beta 1-mRNA abundance increased three- to fourfold at morulation, accompanied by a parallel increase in beta 1-protein synthesis. After midblastula transition (i.e., at early gastrula) and during neurulation, poly(A)+ alpha 1- and beta 3-mRNAs accumulated rapidly, whereas poly(A)+ beta 1-mRNA accumulation was delayed by approximately 2 h, beginning only at early neurula. Our results indicate that 1) the abundance of poly(A)+ beta 1-mRNA is rate limiting during embryonic development for the assembly of alpha 1/beta 1-heterodimers, shown to be involved in the vectorial transport of sodium in kidney cells, and 2) the polyadenylation of beta 1-mRNA is a rate-limiting factor during morulation for the synthesis and assembly of new sodium pumps at the time of blastocoel fluid formation. The 3'-untranslated region of beta 1-mRNA (but not of alpha 1-mRNA) expresses cytoplasmic polyadenylation elements (CPEs) with the consensus sequence AXX-AUUUU(A/U)(A/U)(A/U). A role of CPE in the differential polyadenylation of alpha 1- and beta 1-mRNA is proposed.

摘要

在完全成熟的非洲爪蟾卵母细胞中,β亚基的合成限制了功能性钠钾腺苷三磷酸酶α/β复合物的形成(Geering, K. FEBS Lett. 285: 189 - 193, 1991)。在本研究中,我们发现,在卵母细胞生长过程中(从Ⅰ期到Ⅵ期),α1 - 亚型的mRNA积累,而β1 - 或β3 - 亚型的mRNA不积累。此外,β - mRNA在完全成熟的卵母细胞(Ⅵ期)中显然被隔离在一个未翻译的池中。从受精到桑葚胚形成,α1 - 、β1 - 或β3 - mRNA的总量变化不大。虽然聚腺苷酸化的[poly(A)+]α1 - 和β3 - 亚型mRNA没有显著变化,但在桑葚胚形成时,poly(A)+β1 - mRNA丰度增加了三到四倍,同时β1 - 蛋白合成也相应增加。在囊胚中期转换后(即原肠胚早期)和神经胚形成期间,poly(A)+α1 - 和β3 - mRNA迅速积累,而poly(A)+β1 - mRNA积累延迟约2小时,仅在神经胚早期开始。我们的结果表明:1)在胚胎发育过程中,poly(A)+β1 - mRNA的丰度是α1/β1异二聚体组装的限速因素,α1/β1异二聚体参与肾细胞中钠的向量运输;2)在囊胚腔液形成时,β1 - mRNA的聚腺苷酸化是桑葚胚形成期间新钠泵合成和组装的限速因素。β1 - mRNA(而非α1 - mRNA)的3' - 非翻译区表达具有一致序列AXX - AUUUU(A/U)(A/U)(A/U)的细胞质聚腺苷酸化元件(CPEs)。本文提出了CPE在α1 - 和β1 - mRNA差异聚腺苷酸化中的作用。

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