Genome-wide kinetic properties of transcriptional bursting in mouse embryonic stem cells
Genome-wide kinetic properties of transcriptional bursting in mouse embryonic stem cells
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DOI：
10.1126/sciadv.aaz6699
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发表时间：
2020-06-01
期刊：
影响因子：
13.6
通讯作者：
Nikaido, Itoshi
中科院分区：
文献类型：
--
作者：
Ochiai, Hiroshi;Hayashi, Tetsutaro;Nikaido, Itoshi
转录爆发是启动子的随机激活和失活，导致基因表达的细胞间异质性。然而，哺乳动物细胞中转录爆发动力学（爆发大小和频率）的调节机制仍然难以捉摸。在这项研究中，我们进行了单细胞RNA测序，以分析内在噪音和mRNA水平阐明转录爆发动力学在小鼠胚胎干细胞。信息学分析和功能测定表明，转录爆发动力学调节的启动子和基因体结合蛋白，包括polycomb抑制复合物2和转录延伸因子的组合。此外，基于CRISPR-Cas9的大规模筛选确定Akt/MAPK信号传导途径通过调节转录延伸效率来调节爆发动力学。这些结果揭示了哺乳动物细胞中转录爆发和细胞间基因表达噪音的关键分子机制。
Transcriptional bursting is the stochastic activation and inactivation of promoters, contributing to cell-to-cell heterogeneity in gene expression. However, the mechanism underlying the regulation of transcriptional bursting kinetics (burst size and frequency) in mammalian cells remains elusive. In this study, we performed single-cell RNA sequencing to analyze the intrinsic noise and mRNA levels for elucidating the transcriptional bursting kinetics in mouse embryonic stem cells. Informatics analyses and functional assays revealed that transcriptional bursting kinetics was regulated by a combination of promoter- and gene body-binding proteins, including the polycomb repressive complex 2 and transcription elongation factors. Furthermore, large-scale CRISPR-Cas9-based screening identified that the Akt/MAPK signaling pathway regulated bursting kinetics by modulating transcription elongation efficiency. These results uncovered the key molecular mechanisms underlying transcriptional bursting and cell-to-cell gene expression noise in mammalian cells.