Possible role of mouse poly(A) polymerase mGLD-2 during oocyte maturation
Possible role of mouse poly(A) polymerase mGLD-2 during oocyte maturation
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DOI：
10.1016/j.ydbio.2005.10.017
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发表时间：
2006-01-01
影响因子：
2.7
通讯作者：
Baba, T
中科院分区：
文献类型：
--
作者：
Nakanishi, T;Kubota, H;Baba, T
mRNA的胞质聚腺苷酸化参与基因的转录后调节，包括翻译激活。除了酵母Cid 1和Cid 13以及小鼠TPAP之外，最近已经在秀丽隐杆线虫和Xenoptis卵母细胞中鉴定出GLD-2作为细胞质poly（A）聚合酶。在这项研究中，我们的特点是小鼠GLD-2，mGLD-2，在成年组织，减数分裂成熟的卵母细胞，和NIH 3 T3培养细胞。mGLD-2普遍存在于测试的所有组织和细胞中。mGLD-2定位于体细胞、睾丸细胞和培养细胞的细胞核以及细胞质中。将编码mGLD-2和突变体蛋白的表达质粒转染NIH 3 T3细胞，发现mGLD-2 N端区域的17个残基的序列可能作为转运到细胞核中所需的定位信号。逆转录-聚合酶链反应分析表明，在整个减数分裂成熟的卵母细胞中的mGLD-2 mRNA的存在。然而，54-kDa的mGLD-2被发现在卵母细胞中，只有在中期I和II后，胚泡破裂，大概是由于翻译控制。当mGLD-2的合成，vas人工抑制和双链和多聚腺苷酸化RNA注射到生殖囊泡阶段的卵母细胞，分别增强，卵母细胞成熟显着逮捕中期-I期。这些结果表明，mGLD-2可能在卵母细胞成熟过程中的中期I到中期II的进程中的卵质中发挥作用。(c)2005年爱思唯尔公司All rights reserved.
Cytoplasmic polyadenylation of mRNAs is involved in post-transcriptional regulation of genes, including translational activation. In addition to yeast Cid1 and Cid13 and mouse TPAP, GLD-2 has been recently identified as a cytoplasmic poly(A) polymerase in Caenorhabditis elegans and Xenoptis oocytes. In this study, we have characterized mouse GLD-2, mGLD-2, in adult tissues, meiotically maturing oocytes, and NIH3T3 cultured cells. mGLD-2 was ubiquitously present in all tissues and cells tested. mGLD-2 was localized in the nucleus as well as in the cytoplasm of somatic, testicular, and cultured cells. Transfection of expression plasmids encoding mGLD-2 and the mutant proteins into NIH3T3 cells revealed that a 17-residue sequence in the N-terminal region of mGLD-2 probably acts as a localization signal required for the transport into the nucleus. Analysis of reverse transcriptase-polymerase chain reaction indicated the presence of mGLD-2 mRNA in the oocytes throughout meiotic maturation. However, 54-kDa mGLD-2 was found in the oocytes only at the metaphases I and II after germinal vesicle breakdown, presumably due to translational control. When mGLD-2 synthesis,vas artificially inhibited and enhanced by injection of double-stranded and polyadenylated RNAs into the germinal vesicle-stage oocytes, respectively, oocyte maturation was significantly arrested at the metaphase-I stage. These results suggest that mGLD-2 may act in the ooplasm on the progression of metaphase I to metaphase II during oocyte maturation. (c) 2005 Elsevier Inc. All rights reserved.