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Caenorhabditis elegans Decapping Proteins: Localization and Functional Analysis of Dcp1 Dcp2 and DcpS during Embryogenesis

机译:秀丽隐杆线虫脱壳蛋白:胚胎发生过程中Dcp1Dcp2和DcpS的本地化和功能分析。

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摘要

Though posttranscriptional regulation is important for early embryogenesis, little is understood regarding control of mRNA decay during development. Previous work defined two major pathways by which normal transcripts are degraded in eukaryotes. However it is not known which pathways are key in mRNA decay during early patterning or whether developmental transcripts are turned over via specific pathways. Here we show that Caenorhabditis elegans Dcp2 is localized to distinct foci during embryogenesis, reminiscent of P-bodies, the sites of mRNA degradation in yeast and mammals. However the decapping enzyme of the 3′ to 5′ transcript decay system (DcpS) localizes throughout the cytoplasm, suggesting this degradation pathway is not highly organized. In addition we find that Dcp2 is localized to P-granules, showing that Dcp2 is stored and/or active in these structures. However RNAi of these decapping enzymes has no obvious effect on embryogenesis. In contrast we find that nuclear cap binding proteins (CBP-20 and 80), eIF4G, and PAB-1 are absolutely required for development. Together our data provides further evidence that pathways of general mRNA metabolism can be remarkably organized during development, with two different decapping enzymes localized in distinct cytoplasmic domains.
机译:尽管转录后调控对于早期胚胎发生很重要,但对于控制发育过程中的mRNA衰减了解甚少。先前的工作定义了正常转录本在真核生物中降解的两个主要途径。然而,尚不清楚哪些途径是早期模式中mRNA衰变的关键,还是发育性转录本是否通过特定途径被翻转。在这里,我们显示秀丽隐杆线虫Dcp2在胚胎发生过程中定位于不同的病灶,让人联想到P体,酵母和哺乳动物中mRNA降解的位点。然而,3'到5'转录物衰退系统(DcpS)的去壳酶定位在整个细胞质中,这表明该降解途径的组织性不强。此外,我们发现Dcp2本地化为P颗粒,表明Dcp2在这些结构中存储和/或处于活动状态。然而,这些去壳酶的RNAi对胚胎发生没有明显作用。相比之下,我们发现发育绝对需要核帽结合蛋白(CBP-20和80),eIF4G和PAB-1。我们的数据一起提供了进一步的证据,表明在发育过程中可以显着组织一般的mRNA代谢途径,其中两种不同的去壳酶位于不同的胞质域中。

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