Spatiotemporal coordination of transcription preinitiation complex assembly in live cells.
Spatiotemporal coordination of transcription preinitiation complex assembly in live cells.
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DOI：
10.1016/j.molcel.2021.07.022
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发表时间：
2021-09-02
期刊：
影响因子：
16
通讯作者：
Wu C
中科院分区：
文献类型：
--
作者：
Nguyen VQ;Ranjan A;Liu S;Tang X;Ling YH;Wisniewski J;Mizuguchi G;Li KY;Jou V;Zheng Q;Lavis LD;Lionnet T;Wu C
RNA聚合酶II(Pol II)的转录启动需要在基因启动子上组装预起始复合体(PIC)。在动态核中，数千个启动子广泛分布在染色质中，目前尚不清楚多个单独的成分如何聚集在任何靶点上建立PIC。在这里，我们在酿酒酵母中使用活细胞、单分子跟踪来可视化PIC组件和Mediator在指导这一过程中的关键功能对核质的受限探索。在染色质上，TFIID/TBP、Mediator和POL II指示短暂的PIC的组装，这种组装不经常发生，但平均在几秒钟内有效。此外，核小体侵袭的PIC排除强调了染色质重塑调节启动子的可及性。因此，协调的核探测和对可及靶的招募是在酵母中建立动态PIC的基础。总之，我们的研究为活细胞的转录启动提供了一个全球时空模型。为了表征转录是如何在生理空间和时间中启动的，Nguyen et。艾尔对活酵母中的所有预起始复合体(PIC)组分进行单分子跟踪，揭示它们在染色质上聚合的时空协调，以组装PIC并在几秒钟的时间尺度上启动转录。
Transcription initiation by RNA polymerase II (Pol II) requires preinitiation complex (PIC) assembly at gene promoters. In the dynamic nucleus where thousands of promoters are broadly distributed in chromatin, it is unclear how multiple individual components converge on any target to establish the PIC. Here, we use live-cell, single-molecule tracking in S. cerevisiae to visualize constrained exploration of the nucleoplasm by PIC components and Mediator’s key functions in guiding this process. On chromatin, TFIID/TBP, Mediator, and Pol II instruct assembly of a short-lived PIC, which occurs infrequently but efficiently within a few seconds on average. Moreover, PIC exclusion by nucleosome encroachment underscores regulated promoter accessibility by chromatin remodeling. Thus, coordinated nuclear exploration and recruitment to accessible targets underlies dynamic PIC establishment in yeast. Collectively, our study provides a global spatio-temporal model for transcription initiation in live cells. To characterize how transcription initiates in physiological space and time, Nguyen et. al. perform single-molecule tracking of all preinitiation complex (PIC) components in live yeast, illuminating spatio-temporal coordination underlying their convergence on chromatin to assemble the PIC and initiate transcription on a timescale of several seconds.
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