Akbar Samina, Lee Soon Youl, Boylan Sharon A, Price Chester W
Microbiology (Reading). 1999 May;145 ( Pt 5):1069-1078. doi: 10.1099/13500872-145-5-1069.
The general stress response of Bacillus subtilis is triggered by a variety of environmental and metabolic stresses which activate the sigmaB transcription factor. Among the more than 100 genes controlled by sigmaB (the csb genes), the functions identified thus far include resistance to oxidative stress, resistance to protein denaturation and resistance to osmotic stress. To understand the breadth of functions in which csb genes participate, the transcriptional organization and predicted products of two such genes previously identified in a screen for sigmaB-dependent lacZ fusions were analysed. The csb-22::Tn917lacZ and csb-34::Tn917lacZ fusions are unusual among csb genes in that their expression appears to be completely dependent upon sigmaB. By plasmid-integration experiments, fusion analyses and site-directed mutagenesis, stress-inducible, sigmaB-dependent promoters for both these fusions were identified. The csb-34 fusion marked an ORF (yxcC or csbC) which by sequence analysis lay in a monocistronic transcriptional unit. This ORF encoded a predicted 461-residue product which had high identity with Class I sugar transporters of the major facilitator superfamily. It was speculated that the csbC product could serve either a nutritional or an osmotic protection function. In contrast, the csb-22 fusion identified an ORF (ywmG or csbD) which appeared to be the second gene of a two-gene operon. This ORF encoded a predicted 62-residue product which resembled a small Escherichia coli protein of unknown function. The sigmaB. dependent promoter lay immediately upstream from csbD and appeared to be an internal promoter for the operon.
枯草芽孢杆菌的一般应激反应由多种环境和代谢应激触发,这些应激会激活σB转录因子。在由σB控制的100多个基因(csb基因)中,目前已确定的功能包括对氧化应激的抗性、对蛋白质变性的抗性和对渗透应激的抗性。为了了解csb基因参与的功能广度,分析了之前在筛选σB依赖性lacZ融合体时鉴定的两个此类基因的转录组织和预测产物。csb - 22::Tn917lacZ和csb - 34::Tn917lacZ融合体在csb基因中是不寻常的,因为它们的表达似乎完全依赖于σB。通过质粒整合实验、融合分析和定点诱变,确定了这两个融合体的应激诱导型、σB依赖性启动子。csb - 34融合体标记了一个开放阅读框(yxcC或csbC),通过序列分析,该开放阅读框位于一个单顺反子转录单元中。这个开放阅读框编码了一个预测的461个残基的产物,它与主要易化子超家族的I类糖转运蛋白具有高度同源性。据推测,csbC产物可能具有营养或渗透保护功能。相比之下,csb - 22融合体鉴定出一个开放阅读框(ywmG或csbD),它似乎是一个双基因操纵子的第二个基因。这个开放阅读框编码了一个预测的62个残基的产物,它类似于一种功能未知的小大肠杆菌蛋白。σB依赖性启动子位于csbD的紧邻上游,似乎是该操纵子的内部启动子。