Abuhashem Abderhman, Chivu Alexandra G, Zhao Yixin, Rice Edward J, Siepel Adam, Danko Charles G, Hadjantonakis Anna-Katerina
Developmental Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA.
Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program, New York, New York 10065, USA.
Genes Dev. 2022 Aug 18;36(13-14):770-89. doi: 10.1101/gad.349565.122.
Promoter-proximal RNA Pol II pausing is a critical step in transcriptional control. Pol II pausing has been predominantly studied in tissue culture systems. While Pol II pausing has been shown to be required for mammalian development, the phenotypic and mechanistic details of this requirement are unknown. Here, we found that loss of Pol II pausing stalls pluripotent state transitions within the epiblast of the early mouse embryo. Using mice and a NELFB degron mouse pluripotent stem cell model, we show that embryonic stem cells (ESCs) representing the naïve state of pluripotency successfully initiate a transition program but fail to balance levels of induced and repressed genes and enhancers in the absence of NELF. We found an increase in chromatin-associated NELF during transition from the naïve to later pluripotent states. Overall, our work defines the acute and long-term molecular consequences of NELF loss and reveals a role for Pol II pausing in the pluripotency continuum as a modulator of cell state transitions.
启动子近端RNA聚合酶II（Pol II）暂停是转录控制中的关键步骤。Pol II暂停主要是在组织培养系统中进行研究的。虽然已表明Pol II暂停是哺乳动物发育所必需的，但这一需求的表型和机制细节尚不清楚。在这里，我们发现Pol II暂停的缺失会阻碍小鼠早期胚胎上胚层内多能状态的转变。利用基因敲除小鼠和NELFB降解子小鼠多能干细胞模型，我们表明代表原始多能状态的胚胎干细胞（ESC）成功启动了转变程序，但在缺乏NELF的情况下，无法平衡诱导基因和抑制基因以及增强子的水平。我们发现，从原始多能状态转变为后期多能状态的过程中，与染色质相关的NELF会增加。总体而言，我们的工作定义了NELF缺失的急性和长期分子后果，并揭示了Pol II暂停在多能性连续体中作为细胞状态转变调节剂的作用。
