Essential roles of three enhancer sites in sigma54-dependent transcription by the nitric oxide sensing regulatory protein NorR.

Essential roles of three enhancer sites in sigma54-dependent transcription by the nitric oxide sensing regulatory protein NorR.
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DOI:
10.1093/nar/gkp1065
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发表时间:
2010-03
影响因子:
14.9
通讯作者:
Dixon R
Dixon R
中科院分区:
生物学2区
文献类型:
--
作者:
Tucker NP;Ghosh T;Bush M;Zhang X;Dixon R

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细菌激活蛋白 NorR 与启动子位点上游的增强子样元件结合,并响应 NO 应激,激活编码一氧化氮解毒酶 (NorVW) 的基因的 σ54 依赖性转录。大肠杆菌中norVW启动子的独特之处在于存在与σ54-RNA聚合酶的结合位点相关的三个增强子位点。在这里,我们表明,所有三个位点都是 σ54-RNAP 全酶 (Eσ54) 依赖 NorR 催化开放复合物形成所必需的。我们证明这本质上是由于 NorR 的最大 ATP 酶活性需要所有三种增强子,其中的能量用于重塑封闭的 Eσ54 复合物并允许启动子 DNA 解链。我们还发现位点特异性 DNA 结合本身促进寡聚化,但需要三个位点侧翼的 DNA 来进一步稳定增强子处的 NorR 功能性高阶寡聚物。
The bacterial activator protein NorR binds to enhancer-like elements, upstream of the promoter site, and activates σ54-dependent transcription of genes that encode nitric oxide detoxifying enzymes (NorVW), in response to NO stress. Unique to the norVW promoter in Escherichia coli is the presence of three enhancer sites associated with a binding site for σ54-RNA polymerase. Here we show that all three sites are required for NorR-dependent catalysis of open complex formation by σ54-RNAP holoenzyme (Eσ54). We demonstrate that this is essentially due to the need for all three enhancers for maximal ATPase activity of NorR, energy from which is used to remodel the closed Eσ54 complex and allow melting of the promoter DNA. We also find that site-specific DNA binding per se promotes oligomerisation but the DNA flanking the three sites is needed to further stabilise the functional higher order oligomer of NorR at the enhancers.
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发表时间: 2008-11-07
期刊: MOLECULAR CELL
影响因子: 16
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