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The Flagellar Hook Protein, FlgE, of Salmonella enterica Serovar Typhimurium Is Posttranscriptionally Regulated in Response to the Stage of Flagellar Assembly

机译:肠炎沙门氏菌血清型鼠伤寒的鞭毛钩蛋白FlgE在鞭毛装配阶段响应后转录调控。

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摘要

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.
机译:我们调查了flgE的转录后调控,flgE是编码鞭毛钩亚基蛋白的2类基因。核糖核酸酶保护试验表明,在已知鞭毛组装步骤中有缺陷的许多菌株中,flgE基因以相当的水平转录。但是,对这些菌株进行的Western分析表明FlgE蛋白水平存在实质性差异。尽管在缺失编码开关复合物和鞭毛特异性分泌装置的基因的菌株中观察到了野生型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的水平是否受钩基体组装阶段的影响,对缺失区域内单个位点突变的菌株进行了Western分析。在杆突变体中检测不到FlgE蛋白,在环突变体中检测不到FlgE蛋白,在钩相关蛋白突变体中则检测不到野生型。在杆组装之前,由于开关复合物和鞭毛特异性分泌装置缺失突变体被阻止用于鞭毛构建而缺乏负调控,这表明这些结构在FlgE的负调控中起作用。大量鞭毛突变体的Western定量分析表明FlgE水平反映了鞭毛装配被阻断的阶段。这些数据为响应鞭毛组装阶段的FlgE的转录后负调控提供了证据。

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