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量化活体细胞中 mRNA 向 P 体的靶向性,揭示了它们在 mRNA 降解和储存中的双重作用。

Quantifying mRNA targeting to P-bodies in living human cells reveals their dual role in mRNA decay and storage.

机构信息

The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel.

The Mina & Everard Goodman Faculty of Life Sciences and Institute of Nanotechnology, Bar-Ilan University, Ramat Gan 52900, Israel

出版信息

J Cell Sci. 2014 Oct 15;127(Pt 20):4443-56. doi: 10.1242/jcs.152975. Epub 2014 Aug 15.

Abstract

The 5'-to-3' mRNA degradation machinery localizes to cytoplasmic processing bodies (P-bodies), which are non-membranous structures found in all eukaryotes. Although P-body function has been intensively studied in yeast, less is known about their role in mammalian cells, such as whether P-body enzymes are actively engaged in mRNA degradation or whether P-bodies serve as mRNA storage depots, particularly during cellular stress. We examined the fate of mammalian mRNAs in P-bodies during translational stress, and show that mRNAs accumulate within P-bodies during amino acid starvation. The 5' and 3' ends of the transcripts residing in P-bodies could be identified, but poly(A) tails were not detected. Using the MS2 mRNA-tagging system for mRNA visualization in living cells, we found that a stationary mRNA population formed in P-bodies during translational stress, which cleared gradually after the stress was relieved. Dcp2-knockdown experiments showed that there is constant degradation of part of the P-body-associated mRNA population. This analysis demonstrates the dual role of P-bodies as decay sites and storage areas under regular and stress conditions.

摘要

5'-3' mRNA 降解机制定位于细胞质处理体(P 体),P 体是所有真核生物中发现的非膜结构。尽管 P 体的功能在酵母中已经得到了深入研究,但对于其在哺乳动物细胞中的作用知之甚少,例如 P 体酶是否积极参与 mRNA 降解,或者 P 体是否作为 mRNA 储存库,特别是在细胞应激期间。我们研究了翻译应激期间哺乳动物 mRNA 在 P 体中的命运,并表明在氨基酸饥饿时,mRNA 在 P 体中积累。可以识别 P 体中存在的转录本的 5' 和 3' 末端,但未检测到 poly(A) 尾巴。使用 MS2 mRNA 标记系统在活细胞中可视化 mRNA,我们发现翻译应激期间在 P 体中形成了静止的 mRNA 群体,在应激缓解后逐渐清除。Dcp2 敲低实验表明,P 体相关的 mRNA 群体的一部分持续降解。这项分析表明 P 体在常规和应激条件下具有作为降解部位和储存区域的双重作用。

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