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首页> 外文期刊>Journal of Molecular Biology >Coupling sigma factor conformation to RNA polymerase reorganisation for DNA melting.
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Coupling sigma factor conformation to RNA polymerase reorganisation for DNA melting.

机译：将σ因子构象与RNA聚合酶重组相结合以进行DNA融解。

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摘要

ATP-driven remodelling of initial RNA polymerase (RNAP) promoter complexes occurs as a major post recruitment strategy used to control gene expression. Using a model-enhancer-dependent bacterial system (sigma54-RNAP, Esigma54) and a slowly hydrolysed ATP analogue (ATPgammaS), we provide evidence for a nucleotide-dependent temporal pathway leading to DNA melting involving a small set of sigma54-DNA conformational states. We demonstrate that the ATP hydrolysis-dependent remodelling of Esigma54 occurs in at least two distinct temporal steps. The first detected remodelling phase results in changes in the interactions between the promoter specificity sigma54 factor and the promoter DNA. The second detected remodelling phase causes changes in the relationship between the promoter DNA and the core RNAP catalytic beta/beta' subunits, correlating with the loading of template DNA into the catalytic cleft of RNAP. It would appear that, for Esigma54 promoters, loading of template DNA within the catalytic cleft of RNAP is dependent on fast ATP hydrolysis steps that trigger changes in the beta' jaw domain, thereby allowing acquisition of the open complex status.
机译：ATP驱动的初始RNA聚合酶（RNAP）启动子复合物的重塑是用于控制基因表达的主要后期募集策略。使用模型增强子依赖性细菌系统（sigma54-RNAP，Esigma54）和缓慢水解的ATP类似物（ATPgammaS），我们提供了核苷酸依赖的时间途径的证据，该途径导致DNA融解，涉及一小部分sigma54-DNA构象态。我们证明，Esigma54的ATP水解依赖性重塑发生在至少两个不同的时间步骤中。第一个检测到的重塑阶段导致启动子特异性sigma54因子与启动子DNA之间相互作用的变化。第二个检测到的重塑阶段导致启动子DNA和核心RNAP催化β/β'亚基之间的关系发生变化，这与模板DNA加载到RNAP的催化裂隙中有关。对于Esigma54启动子而言，看来模板DNA在RNA催化裂隙内的负载取决于快速的ATP水解步骤，该步骤会触发β'颚结构域的变化，从而获得开放复合物状态。

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