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首页> 外文期刊>Journal of Molecular Biology >Physical Evidence for the Induced Release of the Bacillus subtilis Transcription Factor, sigma(F), from its Inhibitory Complex.
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Physical Evidence for the Induced Release of the Bacillus subtilis Transcription Factor, sigma(F), from its Inhibitory Complex.

机译：枯草芽孢杆菌转录因子sigma（F）从其抑制复合物中诱导释放的物理证据。

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The release of the transcription factor sigma(F) from its inhibitory complex with SpoIIAB is a key regulatory step in the control of sporulation in Bacillus subtilis as it initiates a pattern of differential gene expression in the mother cell and prespore compartments. The sigma(F).SpoIIAB complex is dissociated by the unphosphorylated form of the protein SpoIIAA, the alternative binding partner of SpoIIAB. Here, we employ fluorescence spectroscopy to examine the mechanism by which SpoIIAA acts on the sigma(F).SpoIIAB complex. We constructed a mutant of sigma(F), sigma(F)-W46L, which displayed a reproducible fluorescence response on binding to SpoIIAB. Using this mutant we were able to quantify the amount of sigma(F) bound to SpoIIAB in real time. The results provide physical evidence for the "induced release" mechanism of sigma(F) activation. We demonstrate that SpoIIAA interacts directly with the sigma(F).SpoIIAB complex, greatly decreasing the affinity of SpoIIAB for sigma(F) and thus causing the release of the latter. We also demonstrate that sigma(F) is released before SpoIIAA is phosphorylated and that release occurs on a similar time scale to the binding of SpoIIAA to SpoIIAB.
机译：从其与SpoIIAB的抑制复合物中释放转录因子sigma（F）是控制枯草芽孢杆菌中孢子形成的关键调控步骤，因为它启动了母细胞和芽孢前区室中差异基因表达的模式。 sigma（F）.SpoIIAB复合物被SpoIIAB的替代结合伴侣SpoIIAA蛋白的非磷酸化形式解离。在这里，我们采用荧光光谱技术来检查SpoIIAA作用于sigma（F）.SpoIIAB复合物的机理。我们构建了一个西格玛（F），西格玛（F）-W46L的突变体，该突变体与SpoIIAB结合后显示出可再现的荧光响应。使用该突变体，我们能够实时定量与SpoIIAB结合的sigma（F）的数量。结果为sigma（F）激活的“诱导释放”机制提供了物理证据。我们证明SpoIIAA直接与sigma（F）.SpoIIAB相互作用，大大降低了SpoIIAB对sigma（F）的亲和力，从而导致后者的释放。我们还证明了在SpoIIAA被磷酸化之前sigma（F）被释放，并且释放发生的时间与SpoIIAA与SpoIIAB的结合时间相似。

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