Records of RNA locations in living yeast revealed through covalent marks.
Records of RNA locations in living yeast revealed through covalent marks.
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DOI：
10.1073/pnas.1921408117
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发表时间：
2020-09-22
影响因子：
11.1
通讯作者：
Wickens M
中科院分区：
文献类型：
--
作者：
Medina-Munoz HC;Lapointe CP;Porter DF;Wickens M
RNA在细胞内移动，通常驻留在特定的位置。在转录组中检测这些运动和位置的需要是紧迫的。我们报告了一种在活细胞中的方法，该方法提供了细胞包含的整个RNA集合的定位记录。它依赖于当RNA直接遇到特定位点时共价标记RNA。使用这种方法，我们确定了一个保守的电池的RNA被确定在一个以上的位置在酵母和人类细胞，非经典模式的本地化，和RNA进行处理或监视在离散的亚细胞位置。我们的研究结果提供了一个入口的历史，个别RNA分子通过共价标记。从胚胎发育到疾病，RNA的移动和定位遍布生物学。为了识别特定位置的RNA，我们开发了一种策略，其中尿苷添加酶锚定到亚细胞位点，在那里它直接用3′末端尿苷标记RNA。这种定位RNA记录方法产生RNA位置的记录，并通过鉴定选择性定位于内质网（ER）或线粒体的RNA进行验证。我们确定了一个广泛的双重定位模式，从酵母到人类细胞，其中相同的电池的mRNA遇到ER和线粒体在这两个物种，包括一个mRNA编码的关键压力传感器。许多多蛋白复合物的亚基定位于ER和线粒体，这表明协调组装。非编码RNA在RNA监视和加工过程中遇到两种细胞器。通过提供随时间推移的RNA位置记录，该方法补充了那些捕获瞬时位置快照的方法。
RNAs move within cells and often reside at specific locations. The need to detect those movements and locales across the transcriptome is pressing. We report an approach in living cells that provides a record of localization across the entire collection of RNAs a cell contains. It relies on covalently marking the RNA when it directly encounters a specific site. Using this approach, we identify a conserved battery of RNAs that are identified at more than one location in yeast and human cells, noncanonical modes of localization, and RNAs undergoing processing or surveillance at discrete subcellular locations. Our findings provide an entrée to the histories of individual RNA molecules through covalent marks. RNA movements and localization pervade biology, from embryonic development to disease. To identify RNAs at specific locations, we developed a strategy in which a uridine-adding enzyme is anchored to subcellular sites, where it directly marks RNAs with 3′ terminal uridines. This localized RNA recording approach yields a record of RNA locations, and is validated through identification of RNAs localized selectively to the endoplasmic reticulum (ER) or mitochondria. We identify a broad dual localization pattern conserved from yeast to human cells, in which the same battery of mRNAs encounter both ER and mitochondria in both species, and include an mRNA encoding a key stress sensor. Subunits of many multiprotein complexes localize to both the ER and mitochondria, suggesting coordinated assembly. Noncoding RNAs in the course of RNA surveillance and processing encounter both organelles. By providing a record of RNA locations over time, the approach complements those that capture snapshots of instantaneous positions.
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