Chen Y F, Helmann J D
Section of Microbiology, Cornell University, Ithaca, NY 14853.
Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5123-7. doi: 10.1073/pnas.89.11.5123.
The activation of additional promoter sites by production of an alternative sigma subunit for RNA polymerase is a common strategy for the coordinate regulation of gene expression. Many alternative sigma factors control genes for specialized, and often narrowly distributed, functions. For example, most of the alternative sigma factors in Bacillus subtilis control genes necessary for endospore formation. In contrast, the B. subtilis sigma D protein controls the expression of genes important for flagellar-based motility and chemotaxis, a form of locomotion very broadly distributed in the eubacteria. A homologous sigma factor, sigma F, controls a similar group of motility genes in the enteric bacteria. The conservation of both promoter specificity and genetic function in these two regulons allowed us to test the ability of a B. subtilis sigma factor to function within an Escherichia coli host. We demonstrate that expression of the B. subtilis sigD gene restores motility to an E. coli strain mutant in the fliA locus encoding the sigma F factor. This result suggests that the B. subtilis sigma D protein can bind to the E. coli core RNA polymerase to direct transcription initiation from at least four of the late operon promoters, thereby leading to the synthesis of flagellin, motor, and hook-associated proteins. Conversely, expression of sigma D protein in a normally chemotactic strain of E. coli (fliA+) leads to a hyperflagellated, nonchemotactic phenotype.
通过产生RNA聚合酶的替代σ亚基来激活额外的启动子位点,是基因表达协调调控的常见策略。许多替代σ因子控制着具有特殊功能且通常分布狭窄的基因。例如,枯草芽孢杆菌中的大多数替代σ因子控制芽孢形成所需的基因。相比之下,枯草芽孢杆菌的σD蛋白控制着对基于鞭毛的运动和趋化性很重要的基因的表达,鞭毛运动是真细菌中广泛分布的一种运动形式。一种同源的σ因子,σF,控制肠道细菌中类似的一组运动基因。这两个调控子中启动子特异性和基因功能的保守性,使我们能够测试枯草芽孢杆菌的一个σ因子在大肠杆菌宿主内发挥功能的能力。我们证明,枯草芽孢杆菌sigD基因的表达能恢复编码σF因子的fliA位点突变的大肠杆菌菌株的运动能力。这一结果表明,枯草芽孢杆菌的σD蛋白可以与大肠杆菌核心RNA聚合酶结合,以指导从至少四个晚期操纵子启动子起始转录,从而导致鞭毛蛋白、马达和钩相关蛋白的合成。相反,在正常具有趋化性的大肠杆菌菌株(fliA+)中表达σD蛋白会导致超鞭毛化、无趋化性的表型。