Downstream sequence-dependent RNA cleavage and pausing by RNA polymerase I
Downstream sequence-dependent RNA cleavage and pausing by RNA polymerase I
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DOI：
10.1074/jbc.ra119.011354
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发表时间：
2020-01-31
影响因子：
4.8
通讯作者：
Schneider, David Alan
中科院分区：
文献类型：
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作者：
Scull, Catherine E.;Clarke, Andrew M.;Schneider, David Alan
DNA模板的序列一直被认为影响DNA依赖性RNA聚合酶的转录速率，但DNA序列对酿酒酵母真核RNA聚合酶I （Pol I）转录伸长特性的影响尚未明确。在这项研究中，我们观察了二核苷酸产生、转录延伸复合物稳定性和Pol I暂停在体外响应下游DNA的变化。体外研究表明，富含at的下游DNA增强了Pol I的暂停作用，抑制了Pol I的核裂解活性。对酿酒酵母Pol I原生延长转录物测序数据的分析表明，这些下游序列元件在体内影响Pol I。原生延长转录本测序研究表明，Pol I占用率随着下游AT含量的增加而增加，而随着下游GC含量的增加而减少。总之，这些数据表明，在体内和体外，下游DNA序列在序列进入Pol I活性位点之前直接影响转录延伸动力学。
The sequence of the DNA template has long been thought to influence the rate of transcription by DNA-dependent RNA polymerases, but the influence of DNA sequence on transcription elongation properties of eukaryotic RNA polymerase I (Pol I) from Saccharomyces cerevisiae has not been defined. In this study, we observe changes in dinucleotide production, transcription elongation complex stability, and Pol I pausing in vitro in response to downstream DNA. In vitro studies demonstrate that AT-rich downstream DNA enhances pausing by Pol I and inhibits Pol I nucleolytic cleavage activity. Analysis of Pol I native elongating transcript sequencing data in Saccharomyces cerevisiae suggests that these downstream sequence elements influence Pol I in vivo. Native elongating transcript sequencing studies reveal that Pol I occupancy increases as downstream AT content increases and decreases as downstream GC content increases. Collectively, these data demonstrate that the downstream DNA sequence directly impacts the kinetics of transcription elongation prior to the sequence entering the active site of Pol I both in vivo and in vitro.