R-loops coordinate with SOX2 in regulating reprogramming to pluripotency
R-loops coordinate with SOX2 in regulating reprogramming to pluripotency
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R 环与 SOX2 协调调节重编程为多能性
DOI：
10.1126/sciadv.aba0777
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发表时间：
2020-06-01
期刊：
影响因子：
13.6
通讯作者：
Yao, Hongjie
中科院分区：
文献类型：
--
作者：
Li, Yaoyi;Song, Yawei;Yao, Hongjie
R 环动态发生在基因表达变化之前，并且是体细胞重编程所必需的。 R环调节基因组稳定性并调节基因表达，但R环在干细胞生物学中的功能和调节机制仍不清楚。在这里，我们分析了体细胞重编程过程中的 R 环，发现 R 环的动态变化对于重编程至关重要，并且发生在基因表达变化之前。通过耗尽 RNaseH1 或在 D209 (RNaseH1D209N) 处催化失活 RNaseH1 来破坏 R 环的稳态会阻止重编程。 Sox2（但不是山中鸡尾酒中的任何其他因子）克服了 RNaseH1 活性丧失对重编程的抑制作用。 Sox2 与重编程屏障因子 Ddx5 相互作用，抑制 R 环上 Ddx5 的解离酶活性，从而促进重编程。此外，重编程效率可以通过 dCas9 介导的针对特定 R 环区域的 RNaseH1/RNaseH1D209N 进行调节。总之，这些结果表明 R 环在重编程中发挥重要作用，并揭示了重编程过程中 Sox2/Ddx5 对 R 环的调节模块。
R-loop dynamics occur before changes in gene expression and are required for somatic cell reprogramming. R-loops modulate genome stability and regulate gene expression, but the functions and the regulatory mechanisms of R-loops in stem cell biology are still unclear. Here, we profiled R-loops during somatic cell reprogramming and found that dynamic changes in R-loops are essential for reprogramming and occurred before changes in gene expression. Disrupting the homeostasis of R-loops by depleting RNaseH1 or catalytic inactivation of RNaseH1 at D209 (RNaseH1D209N) blocks reprogramming. Sox2, but not any other factor in the Yamanaka cocktail, overcomes the inhibitory effects of RNaseH1 activity loss on reprogramming. Sox2 interacts with the reprogramming barrier factor Ddx5 and inhibits the resolvase activity of Ddx5 on R-loops and thus facilitates reprogramming. Furthermore, reprogramming efficiency can be modulated by dCas9-mediated RNaseH1/RNaseH1D209N targeting the specific R-loop regions. Together, these results show that R-loops play important roles in reprogramming and shed light on the regulatory module of Sox2/Ddx5 on R-loops during reprogramming.