Defining the status of RNA polymerase at promoters.
Defining the status of RNA polymerase at promoters.
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DOI：
10.1016/j.celrep.2012.08.034
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发表时间：
2012-10-25
期刊：
影响因子：
8.8
通讯作者：
Lis JT
中科院分区：
文献类型：
--
作者：
Core LJ;Waterfall JJ;Gilchrist DA;Fargo DC;Kwak H;Adelman K;Lis JT
最近对后生动物的全基因组研究表明，RNA聚合酶II(POL II)在转录过程中以限速步骤在许多启动子上积累到高密度。然而，这一POL II的地位仍然是一个有争议的领域。在这里，我们比较了GRO-SEQ和CHIP-SEQ分析的定量结果，并证明了果蝇启动子上的大部分POL II是转录参与的-非常少存在于预启动或被阻止的复合体中。这些启动子-近端聚合酶被洗涤剂敏感因子抑制进一步伸长，而负伸长因子NELF的敲除降低了它们的水平。这些结果不仅证实了停顿发生在大多数启动子中，而且证明了停顿是这些启动子早期转录的主要限速步骤。最后，在哺乳动物启动子中看到的发散的延长复合体在果蝇中的比例要低得多，这种定向特异性与果蝇中丰富的定向核心启动子元件有关。
Recent genome-wide studies in metazoans have shown that RNA Polymerase II (Pol II) accumulates to high densities on many promoters at a rate-limited step in transcription. However, the status of this Pol II remains an area of debate. Here, we compare quantitative outputs of GRO-seq and ChIP-seq assays and demonstrate the majority of the Pol II on Drosophila promoters is transcriptionally-engaged - very little exists in a preinitiation or arrested complex. These promoter-proximal polymerases are inhibited from further elongation by detergent sensitive factors, and knockdown of negative elongation factor, NELF, reduces their levels. These results not only solidify that pausing occurs at most promoters, but demonstrate that it is the major rate-limiting step in early transcription at these promoters. Finally, the divergent elongation complexes seen at mammalian promoters are far less prevalent in Drosophila, and this specificity in orientation correlates with directional core promoter elements, which are abundant in Drosophila.
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1073/pnas.0910177106
发表时间：
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