Super resolution microscopy reveals how elongating RNA polymerase II and nascent RNA interact with nucleosome clutches.
Super resolution microscopy reveals how elongating RNA polymerase II and nascent RNA interact with nucleosome clutches.
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DOI：
10.1093/nar/gkab1215
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发表时间：
2022-01-11
影响因子：
14.9
通讯作者：
Cosma MP
中科院分区：
文献类型：
--
作者：
Castells-Garcia A;Ed-Daoui I;González-Almela E;Vicario C;Ottestrom J;Lakadamyali M;Neguembor MV;Cosma MP
转录和基因组结构是相互依赖的，但目前还不清楚染色质纤维中的核小体如何与新生RNA相互作用，以及这些RNA和延伸RNA聚合酶II（RNAP II）的相对核分布。使用超分辨率（SR）显微镜，我们可视化的新生转录组，在核质和核仁，与纳米级的分辨率。我们发现，新生的RNA组织在我们称为RNA纳米结构域的结构中，其特征与随时间产生的转录本数量无关。新生RNA的双色SR成像，以及延长RNAP II和H2B，显示了核小体离合器，RNAP II和RNA纳米结构域之间的物理关系。核小体离合器和RNA纳米结构域之间的距离大于延长RNAP II和RNA纳米结构域之间测量的距离。延伸RNAP II位于新生RNA和小的转录活性核小体离合器之间。此外，RNA工厂很小，主要由少数RNAP II形成。最后，我们描述了一种新的方法来量化在一个单独的基因位点的转录活性。通过测量单个等位基因转录激活后的局部新生RNA积累，我们证实了在全球核水平上进行的测量。描绘核小体离合器、RNAP II簇和RNA纳米结构域之间的关联的示意图。染色质（绿色）以离合器的形式排列。小的活性核小体离合器与伸长的RNAP II簇（紫色）相关，RNAP II簇产生小的RNA纳米结构域（红色）。
Transcription and genome architecture are interdependent, but it is still unclear how nucleosomes in the chromatin fiber interact with nascent RNA, and which is the relative nuclear distribution of these RNAs and elongating RNA polymerase II (RNAP II). Using super-resolution (SR) microscopy, we visualized the nascent transcriptome, in both nucleoplasm and nucleolus, with nanoscale resolution. We found that nascent RNAs organize in structures we termed RNA nanodomains, whose characteristics are independent of the number of transcripts produced over time. Dual-color SR imaging of nascent RNAs, together with elongating RNAP II and H2B, shows the physical relation between nucleosome clutches, RNAP II, and RNA nanodomains. The distance between nucleosome clutches and RNA nanodomains is larger than the distance measured between elongating RNAP II and RNA nanodomains. Elongating RNAP II stands between nascent RNAs and the small, transcriptionally active, nucleosome clutches. Moreover, RNA factories are small and largely formed by few RNAP II. Finally, we describe a novel approach to quantify the transcriptional activity at an individual gene locus. By measuring local nascent RNA accumulation upon transcriptional activation at single alleles, we confirm the measurements made at the global nuclear level. Schematic representation depicting the association between nucleosome clutches, RNAP II clusters and RNA nanodomains. Chromatin (in green) is organized in clutches. Small active nucleosome clutches are associated with elongating RNAP II clusters (in purple) which generates small RNA nanodomains (in red).
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影响因子：
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