Transcription activation at promoters carrying tandem DNA sites for the Escherichia coli cyclic AMP receptor protein:: Organisation of the RNA polymerase α subunits
Transcription activation at promoters carrying tandem DNA sites for the Escherichia coli cyclic AMP receptor protein:: Organisation of the RNA polymerase α subunits
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DOI：
10.1006/jmbi.1998.1666
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发表时间：
1998-04-10
影响因子：
5.6
通讯作者：
Busby, SJW
中科院分区：
文献类型：
--
作者：
Belyaeva, TA;Rhodius, VA;Busby, SJW
我们构建了一系列携带大肠杆菌环AMP受体蛋白（CRP）串联DNA位点的启动子家族，其中一个位点位于转录起始位点上游碱基对41和42之间，第二个位点位于更上游。体内活性测量表明，这些启动子的活性完全依赖于 CRP，并且根据精确位置，结合在上游位点的 CRP 会增加转录激活。利用羟基自由基足迹来研究 CRP 和 RNA 聚合酶全酶 (RNAP) 与这些启动子的结合。研究表明，RNAP α 亚基的 C 末端结构域与上游 CRP 相邻结合，并且它们的精确定位取决于上游结合 CRP 的位置。 RNAP α 亚基的 C 末端结构域通过激活 CRP 的区域 1 与上游和下游结合的 CRP 相互作用。 (C) 1998 学术出版社有限公司。
We have constructed a family of promoters carrying tandem DNA sites for the Escherichia coli cyclic AMP receptor protein (CRP), with one of the sites centred between base-pairs 41 and 42 upstream from the transcription start site, and the second site located further upstream. In vivo activity measurements show that the activity of these promoters is completely dependent on CRP and that, depending on the precise location, CRP bound at the upstream site increases transcription activation. Hydroxyl radical footprinting was exploited to investigate the binding of CRP and RNA polymerase holoenzyme (RNAP) to these promoters. The study shows that the C-terminal domains of the RNAP alpha subunits bind adjacent to the upstream CRP and that their Precise positioning depends on the location of upstream-bound CRP. The C-terminal domains of the RNAP alpha subunits interact with both the upstream and downstream-bound CRP via activating region 1 of CRP. (C) 1998 Academic Press Limited .