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鼠伤寒沙门氏菌的鞭毛钩蛋白FlgE在转录后受到调控,以响应鞭毛组装的阶段。

The flagellar hook protein, FlgE, of Salmonella enterica serovar typhimurium is posttranscriptionally regulated in response to the stage of flagellar assembly.

作者信息

Bonifield H R, Yamaguchi S, Hughes K T

机构信息

Department of Microbiology, University of Washington, Seattle 98195, USA.

出版信息

J Bacteriol. 2000 Jul;182(14):4044-50. doi: 10.1128/JB.182.14.4044-4050.2000.

Abstract

We investigated the posttranscriptional regulation of flgE, a class 2 gene that encodes the hook subunit protein of the flagella. RNase protection assays demonstrated that the flgE gene was transcribed at comparable levels in numerous strains defective in known steps of flagellar assembly. However, Western analyses of these strains demonstrated substantial differences in FlgE protein levels. Although wild-type FlgE levels were observed in strains with deletions of genes encoding components of the switch complex and the flagellum-specific secretion apparatus, no protein was detected in a strain with deletions of the rod, ring, and hook-associated proteins. To determine whether FlgE levels were affected by the stage of hook-basal-body assembly, Western analysis was performed on strains with mutations at individual loci encompassed by the deletion. FlgE protein was undetectable in rod mutants, intermediate in ring mutants, and wild type in hook-associated protein mutants. The lack of negative regulation in switch complex and flagellum-specific secretion apparatus deletion mutants blocked for flagellar construction prior to rod assembly suggests that these structures play a role in the negative regulation of FlgE. Quantitative Western analyses of numerous flagellar mutants indicate that FlgE levels reflect the stage at which flagellar assembly is blocked. These data provide evidence for negative posttranscriptional regulation of FlgE in response to the stage of flagellar assembly.

摘要

我们研究了flgE的转录后调控,flgE是一个2类基因,编码鞭毛的钩亚基蛋白。核糖核酸酶保护分析表明,flgE基因在许多鞭毛组装已知步骤存在缺陷的菌株中以相当的水平转录。然而,对这些菌株的蛋白质免疫印迹分析表明,FlgE蛋白水平存在显著差异。虽然在缺失编码开关复合体和鞭毛特异性分泌装置组件的基因的菌株中观察到野生型FlgE水平,但在缺失杆、环和钩相关蛋白的菌株中未检测到蛋白质。为了确定FlgE水平是否受钩基体组装阶段的影响,对缺失所涵盖的各个位点发生突变的菌株进行了蛋白质免疫印迹分析。在杆突变体中未检测到FlgE蛋白,在环突变体中为中等水平,而在钩相关蛋白突变体中为野生型水平。在杆组装之前被阻断的鞭毛构建的开关复合体和鞭毛特异性分泌装置缺失突变体中缺乏负调控,这表明这些结构在FlgE的负调控中起作用。对众多鞭毛突变体的定量蛋白质免疫印迹分析表明,FlgE水平反映了鞭毛组装被阻断的阶段。这些数据为FlgE在转录后水平上响应鞭毛组装阶段的负调控提供了证据。

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