Repeat RNAs associate with replication forks and post-replicative DNA
Repeat RNAs associate with replication forks and post-replicative DNA
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DOI：
10.1261/rna.074757.120
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发表时间：
2020-09-01
期刊：
影响因子：
4.5
通讯作者：
Lund, Anders H.
中科院分区：
文献类型：
--
作者：
Gylling, Helene M.;Gonzalez-Aguilera, Cristina;Lund, Anders H.
非编码RNA作为DNA和组蛋白修饰复合物之间的支架，具有指导和调节染色质修饰的能力。然而，目前尚不清楚ncRNA是否在DNA复制和表观基因组维持中发挥任何作用，包括组蛋白驱逐和基因组复制后组蛋白修饰的重新安置。新生染色质的分离已经确定了大量的RNA结合蛋白，除了未知的复制和表观遗传维持机制的组成部分。在这里，我们分离和特征化的长和短RNA与新生的染色质在活跃的复制叉和跟踪RNA的组成，在整个细胞周期的染色质成熟。在分叉通道后不久，富含GA的RNA、含有α的RNA和含有TElomeric Repeat-containing RNA（TERRA）与复制的DNA相关。这些含有重复序列的RNA来自进行复制的基因座，表明顺式相互作用。在染色质成熟期间的复制后，甚至在G1中的有丝分裂后，重复序列仍然富集在DNA上。这表明特定类型的重复RNA在DNA复制后不久就被转录，并在整个细胞周期中与其起源基因座稳定结合。所提出的方法和数据使得能够研究RNA与复制叉和复制后染色质的相互作用，并提供关于DNA复制和跨细胞周期如何调节重复RNA及其与染色质的接合的见解。
Noncoding RNA has a proven ability to direct and regulate chromatin modifications by acting as scaffolds between DNA and histone-modifying complexes. However, it is unknown if ncRNA plays any role in DNA replication and epigenome maintenance, including histone eviction and reinstallment of histone modifications after genome duplication. Isolation of nascent chromatin has identified a large number of RNA-binding proteins in addition to unknown components of the replication and epigenetic maintenance machinery. Here, we isolated and characterized long and short RNAs associated with nascent chromatin at active replication forks and track RNA composition during chromatin maturation across the cell cycle. Shortly after fork passage, GA-rich-, alpha- and TElomeric Repeat-containing RNAs (TERRA) are associated with replicated DNA. These repeat containing RNAs arise from loci undergoing replication, suggesting an interaction in cis. Post-replication during chromatin maturation, and even after mitosis in G1, the repeats remain enriched on DNA. This suggests that specific types of repeat RNAs are transcribed shortly after DNA replication and stably associate with their loci of origin throughout the cell cycle. The presented method and data enable studies of RNA interactions with replication forks and post-replicative chromatin and provide insights into how repeat RNAs and their engagement with chromatin are regulated with respect to DNA replication and across the cell cycle.