Transient translational quiescence in primordial germ cells
Transient translational quiescence in primordial germ cells
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DOI：
10.1242/dev.144170
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发表时间：
2017-04-01
期刊：
影响因子：
4.6
通讯作者：
Wessel, Gary M.
中科院分区：
文献类型：
--
作者：
Oulhen, Nathalie;Swartz, S. Zachary;Wessel, Gary M.
动物中的干细胞通常表现出缓慢的细胞周期和/或称为静止的低转录活性。在此，我们报道了海胆胚胎(Strongyloctus PurPuratus)原始生殖细胞(PGCs)中的翻译活性是静止的。我们用O-丙叉嘌呤和L-丙叉甘氨酸Click-it技术测量了新的蛋白质合成，并确定这些细胞合成蛋白质的水平仅为相邻体细胞的6%。被吗啉反义寡核苷酸抑制RNA结合蛋白Nanos2的翻译，或由CRISPR/Cas9敲除Nanos2基因，导致一般翻译显著但部分增加(47%)，尤其是在PGCs。我们发现翻译因子eEF1a的mRNA以一种Nanos2依赖的方式从PGCs中排除在外，这是其3‘UTR中的Nanos/Pumilio反应元件(Pre)的结果。除了eEF1a外，PGCs的胞浆pH似乎抑制翻译，简单地增加pH也能显著选择性地恢复PGCs的翻译。我们得出的结论是，这种海胆的PGC建立了平行的路径，以彻底但短暂地停止翻译。
Stem cells in animals often exhibit a slow cell cycle and/or low transcriptional activity referred to as quiescence. Here, we report that the translational activity in the primordial germ cells (PGCs) of the sea urchin embryo (Strongylocentrotus purpuratus) is quiescent. We measured new protein synthesis with O-propargyl-puromycin and Lhomopropargylglycine Click-iT technologies, and determined that these cells synthesize protein at only 6% the level of their adjacent somatic cells. Knockdown of translation of the RNA-binding protein Nanos2 by morpholino antisense oligonucleotides, or knockout of the Nanos2 gene by CRISPR/Cas9 resulted in a significant, but partial, increase (47%) in general translation specifically in the PGCs. We found that the mRNA of the translation factor eEF1A is excluded from the PGCs in a Nanos2-dependent manner, a consequence of a Nanos/Pumilio response element (PRE) in its 3 ' UTR. In addition to eEF1A, the cytoplasmic pH of the PGCs appears to repress translation and simply increasing the pH also significantly restores translation selectively in the PGCs. We conclude that the PGCs of this sea urchin institute parallel pathways to quiesce translation thoroughly but transiently.