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单核细胞增生李斯特菌中一般应激σ因子σB对相容性溶质转运蛋白转录的调控。

Regulation of transcription of compatible solute transporters by the general stress sigma factor, sigmaB, in Listeria monocytogenes.

作者信息

Cetin Mehmet Sevket, Zhang Chaomei, Hutkins Robert W, Benson Andrew K

机构信息

Department of Food Science and Technology, University of Nebraska, Lincoln, Nebraska 68583-0919, USA.

出版信息

J Bacteriol. 2004 Feb;186(3):794-802. doi: 10.1128/JB.186.3.794-802.2004.

Abstract

Listeria monocytogenes is well known for its durable physiological characteristics, which allow the organism to grow at low temperature and pH and high osmolarity. Growth under high osmolarity depends on the accumulation of compatible solutes, among which glycine betaine and carnitine are the preferred solutes for this organism. Three different transport systems, Gbu, BetL, and OpuC, have been identified in L. monocytogenes which serve to scavenge the preferred compatible solutes. The general stress response regulator sigma(B) has been shown to play an important role in osmotic adaptation in L. monocytogenes, presumably by directing transcription from one or more of the solute transport genes. In the studies presented here, we have used primer extension analyses to identify the promoter elements responsible for transcription of the opuC, gbuA, and betL genes. All three genes are osmotically inducible to some degree. betL is transcribed from a sigma(B)-independent promoter, while gbuA is transcribed from dual promoters, one of which is sigma(B) dependent. opuC is transcribed exclusively from a sigma(B)-dependent promoter. The betL promoter is similar in sequence to the sigma(B)-independent gbuAP1 promoter. Kinetic analysis of transcript accumulation after osmotic upshift demonstrated that sigma(B)-dependent transcripts from gbuAP2 and sigB accumulate for an extended period after upshift, suggesting that sigma(B) activity may provide a mechanism for sustained high-level expression during osmotic challenge. In contrast to osmotic upshift, expression from the sigma(B)-dependent opuC and gbuAP2 promoters after temperature upshift and ethanol stress was minimal, suggesting that additional mechanisms may also participate in regulating transcription from these sigma(B)-dependent promoters.

摘要

单核细胞增生李斯特菌以其持久的生理特性而闻名,这些特性使该生物体能够在低温、低pH值和高渗透压条件下生长。在高渗透压下生长依赖于相容性溶质的积累,其中甘氨酸甜菜碱和肉碱是该生物体的首选溶质。在单核细胞增生李斯特菌中已鉴定出三种不同的转运系统,即Gbu、BetL和OpuC,它们用于清除首选的相容性溶质。一般应激反应调节因子sigma(B)已被证明在单核细胞增生李斯特菌的渗透适应中起重要作用,大概是通过指导一个或多个溶质转运基因的转录来实现的。在本文所呈现的研究中,我们使用引物延伸分析来鉴定负责opuC、gbuA和betL基因转录的启动子元件。这三个基因在某种程度上都受到渗透压诱导。betL从一个不依赖sigma(B)的启动子转录,而gbuA从双重启动子转录,其中一个是依赖sigma(B)的。opuC仅从一个依赖sigma(B)的启动子转录。betL启动子在序列上与不依赖sigma(B)的gbuAP1启动子相似。渗透压上调后转录本积累的动力学分析表明,gbuAP2和sigB的依赖sigma(B)的转录本在上调后会持续积累一段时间,这表明sigma(B)活性可能为渗透挑战期间的持续高水平表达提供一种机制。与渗透压上调相反,温度上调和乙醇应激后依赖sigma(B)的opuC和gbuAP2启动子的表达极少,这表明可能还有其他机制参与调节这些依赖sigma(B)的启动子的转录。

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