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鼠伤寒沙门氏菌中fis的序列、调控及功能

Sequence, regulation, and functions of fis in Salmonella typhimurium.

作者信息

Osuna R, Lienau D, Hughes K T, Johnson R C

机构信息

Department of Biological Chemistry, School of Medicine, University of California, Los Angeles 90024, USA.

出版信息

J Bacteriol. 1995 Apr;177(8):2021-32. doi: 10.1128/jb.177.8.2021-2032.1995.

Abstract

The fis operon from Salmonella typhimurium has been cloned and sequenced, and the properties of Fis-deficient and Fis-constitutive strains were examined. The overall fis operon organization in S. typhimurium is the same as that in Escherichia coli, with the deduced Fis amino acid sequences being identical between both species. While the open reading frames upstream of fis have diverged slightly, the promoter regions between the two species are also identical between -49 and +94. Fis protein and mRNA levels fluctuated dramatically during the course of growth in batch cultures, peaking at approximately 40,000 dimers per cell in early exponential phase, and were undetectable after growth in stationary phase. fis autoregulation was less effective in S. typhimurium than that in E. coli, which can be correlated with the absence or reduced affinity of several Fis-binding sites in the S. typhimurium fis promoter region. Phenotypes of fis mutants include loss of Hin-mediated DNA inversion, cell filamentation, reduced growth rates in rich medium, and increased lag times when the mutants are subcultured after prolonged growth in stationary phase. On the other hand, cells constitutively expressing Fis exhibited normal logarithmic growth but showed a sharp reduction in survival during stationary phase. During the course of these studies, the sigma 28-dependent promoter within the hin-invertible segment that is responsible for fljB (H2) flagellin synthesis was precisely located.

摘要

鼠伤寒沙门氏菌的fis操纵子已被克隆和测序,并对Fis缺陷型和Fis组成型菌株的特性进行了研究。鼠伤寒沙门氏菌中fis操纵子的整体组织与大肠杆菌中的相同,两种物种推导的Fis氨基酸序列相同。虽然fis上游的开放阅读框略有差异,但两个物种之间的启动子区域在-49至+94之间也相同。在分批培养的生长过程中,Fis蛋白和mRNA水平剧烈波动,在指数早期阶段达到约每细胞40,000个二聚体的峰值,在稳定期生长后无法检测到。fis自调控在鼠伤寒沙门氏菌中比在大肠杆菌中效果更差,这可能与鼠伤寒沙门氏菌fis启动子区域中几个Fis结合位点的缺失或亲和力降低有关。fis突变体的表型包括Hin介导的DNA倒位丧失、细胞丝状化、在丰富培养基中生长速率降低以及在稳定期长时间生长后传代培养时延迟期增加。另一方面,组成型表达Fis的细胞在对数期生长正常,但在稳定期存活率急剧下降。在这些研究过程中,精确确定了负责fljB(H2)鞭毛蛋白合成的hin可逆片段内依赖于sigma 28的启动子。

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