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The core-independent promoter-specific binding of Bacillus subtilis sigma(B)

机译:枯草芽孢杆菌西格玛(B)的非核心启动子特异性结合

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摘要

Bacillussubtilis sigma(D) is an alternative sigma factor that possesses a core-independent promoter -10 element binding specificity despite the lack of a distinct footprint on its cognate promoter. We wished to determine whether this property is common to alternative sigma factors. To this end, we over-expressed B.subtilis sigma(B) in Escherichiacoli and analyzed its DNA binding ability by electrophoretic mobility shift assay and DNaseI footprinting. The major complex formed by sigma(B) and its cognate promoter DNA is heparin-sensitive. However, in contrast to the -10 element binding specificity observed for B.subtilis sigma(D), the promoter binding of sigma(B) is specific for the -35 element. These and other results clearly demonstrate that alternative sigma factors possess different promoter-binding characteristics, and make core-independent contributions to recognition of their cognate promoters.
机译:枯草芽孢杆菌sigma(D)是另一种sigma因子,尽管其同源启动子上没有明显的印迹,但它具有不依赖核心的启动子-10元素结合特异性。我们希望确定此属性对于替代的sigma因子是否通用。为此,我们在大肠杆菌中过量表达了枯草芽孢杆菌sigma(B),并通过电泳迁移率测定和DNaseI足迹分析了其DNA结合能力。由sigma(B)及其同源启动子DNA形成的主要复合物对肝素敏感。但是,与枯草芽孢杆菌sigma(D)观察到的-10个元素结合特异性相反,sigma(B)的启动子结合对-35元素具有特异性。这些和其他结果清楚地表明,替代的sigma因子具有不同的启动子结合特征,并为识别其同源启动子做出了与核无关的贡献。

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