Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom.
Department of Computing, Imperial College London, London, United Kingdom.
PLoS Biol. 2023 Jan 17;21(1):e3001956. doi: 10.1371/journal.pbio.3001956. eCollection 2023 Jan.
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.
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。