Kinetics of promoter Pol II on Hsp70 reveal stable pausing and key insights into its regulation.
Kinetics of promoter Pol II on Hsp70 reveal stable pausing and key insights into its regulation.
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DOI：
10.1101/gad.231886.113
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发表时间：
2014-01-01
影响因子：
10.5
通讯作者：
Lis JT
中科院分区：
文献类型：
--
作者：
Buckley MS;Kwak H;Zipfel WR;Lis JT
动力学与启动子近端暂停Pol II经历提前终止与生产性延长是关键的了解后生动物转录调控机制。为了评估暂停的Pol II的命运，Lis及其同事通过光学研究和生化分析的结合测量了暂停的Pol II在果蝇Hsp70位点的稳定性。热休克大大增加延长Pol II而不减少终止，表明调节作用在暂停Pol II进入生产性伸长的步骤。启动子近端暂停的RNA聚合酶II（Pol II）经历过早终止与生产延长的动力学是理解后生动物转录调控的基本机制的核心。为了评估命运的Pol II定量，我们跟踪光活化GFP标记的Pol II在未诱导的热休克蛋白70多线染色体，并表明，Pol II是稳定暂停与半衰期为5分钟。短新生RNA从热休克蛋白70的生化分析表明，这个半衰期是由两个可比的生产延长率和提前终止暂停Pol II。重要的是，热休克显着增加延长Pol II而不减少终止，表明调节作用在暂停Pol II进入生产性伸长的步骤。
The kinetics with which promoter-proximal paused Pol II undergoes premature termination versus productive elongation is key to understanding mechanisms of metazoan transcription regulation. To assess the fate of paused Pol II, Lis and colleagues measured the stability of paused Pol II at the Drosophila Hsp70 locus through a combination of optical studies and biochemical analysis. Heat shock drastically increases elongating Pol II without decreasing termination, indicating that regulation acts at the step of paused Pol II entry to productive elongation. The kinetics with which promoter-proximal paused RNA polymerase II (Pol II) undergoes premature termination versus productive elongation is central to understanding underlying mechanisms of metazoan transcription regulation. To assess the fate of Pol II quantitatively, we tracked photoactivatable GFP-tagged Pol II at uninduced Hsp70 on polytene chromosomes and showed that Pol II is stably paused with a half-life of 5 min. Biochemical analysis of short nascent RNA from Hsp70 reveals that this half-life is determined by two comparable rates of productive elongation and premature termination of paused Pol II. Importantly, heat shock dramatically increases elongating Pol II without decreasing termination, indicating that regulation acts at the step of paused Pol II entry to productive elongation.
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影响因子：
8.8
作者：
Core LJ;Waterfall JJ;Gilchrist DA;Fargo DC;Kwak H;Adelman K;Lis JT
通讯作者：
Lis JT
影响因子：
16
作者：
Anindya, Roy;Ayguen, Ozan;Svejstrup, Jesper Q.
通讯作者：
Svejstrup, Jesper Q.
影响因子：
16
作者：
Saeki, Hideaki;Svejstrup, Jesper Q.
通讯作者：
Svejstrup, Jesper Q.
DOI：
10.1073/pnas.90.17.7923
发表时间：
1993-09-01
影响因子：
11.1
作者：
RASMUSSEN, EB;LIS, JT
通讯作者：
LIS, JT
影响因子：
30.8
作者：
Zeitlinger, Julia;Stark, Alexander;Young, Richard A.
通讯作者：
Young, Richard A.