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在果蝇胚胎中结合 mRNA 衰减动力学建模和单分子成像揭示了 P 体在 5' 到 3' 降解中的作用。

Combined modelling of mRNA decay dynamics and single-molecule imaging in the Drosophila embryo uncovers a role for P-bodies in 5' to 3' degradation.

机构信息

Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom.

Department of Computing, Imperial College London, London, United Kingdom.

出版信息

PLoS Biol. 2023 Jan 17;21(1):e3001956. doi: 10.1371/journal.pbio.3001956. eCollection 2023 Jan.

Abstract

Regulation of mRNA degradation is critical for a diverse array of cellular processes and developmental cell fate decisions. Many methods for determining mRNA half-lives rely on transcriptional inhibition or metabolic labelling. Here, we use a non-invasive method for estimating half-lives for hundreds of mRNAs in the early Drosophila embryo. This approach uses the intronic and exonic reads from a total RNA-seq time series and Gaussian process regression to model the dynamics of premature and mature mRNAs. We show how regulation of mRNA stability is used to establish a range of mature mRNA dynamics during embryogenesis, despite shared transcription profiles. Using single-molecule imaging, we provide evidence that, for the mRNAs tested, there is a correlation between short half-life and mRNA association with P-bodies. Moreover, we detect an enrichment of mRNA 3' ends in P-bodies in the early embryo, consistent with 5' to 3' degradation occurring in P-bodies for at least a subset of mRNAs. We discuss our findings in relation to recently published data suggesting that the primary function of P-bodies in other biological contexts is mRNA storage.

摘要

mRNA 降解的调控对于多种细胞过程和发育细胞命运决定至关重要。许多确定 mRNA 半衰期的方法都依赖于转录抑制或代谢标记。在这里,我们使用一种非侵入性方法来估计早期果蝇胚胎中数百个 mRNA 的半衰期。这种方法利用来自总 RNA-seq 时间序列的内含子和外显子读数,以及高斯过程回归来模拟不成熟和成熟 mRNA 的动态。我们展示了尽管转录谱共享,但如何利用 mRNA 稳定性的调节来在胚胎发生过程中建立一系列成熟 mRNA 的动态。通过单分子成像,我们提供了证据表明,对于测试的 mRNAs,半衰期短与 mRNA 与 P 体的关联之间存在相关性。此外,我们在早期胚胎中检测到 P 体中 mRNA 3' 末端的富集,这与至少一部分 mRNAs 的 5' 到 3' 降解发生在 P 体中一致。我们将我们的发现与最近发表的数据进行了讨论,这些数据表明 P 体在其他生物背景下的主要功能是 mRNA 储存。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/172b/9882958/b5f1b32fbdd3/pbio.3001956.g001.jpg

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