Muir Rachel E, Gober James W
Department of Chemistry and Biochemistry and Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095-1569, USA.
J Bacteriol. 2005 Feb;187(3):949-60. doi: 10.1128/JB.187.3.949-960.2005.
In the Caulobacter crescentus predivisional cell, class III and IV flagellar genes, encoding the extracytoplasmic components of the flagellum, are transcribed in the nascent swarmer compartment. This asymmetric expression pattern is attributable to the compartmentalized activity of the sigma54-dependent transcriptional activator FlbD. Additionally, these temporally transcribed flagellar promoters possess a consensus sequence for the DNA-binding protein integration host factor (IHF), located between the upstream FlbD binding site and the promoter sequences. Here, we deleted the C. crescentus gene encoding the beta-subunit of the IHF, ihfB (himD), and examined the effect on flagellar gene expression. The DeltaihfB strain exhibited a mild defect in cell morphology and impaired motility. Using flagellar promoter reporter fusions, we observed that expression levels of a subset of class III flagellar promoters were decreased by the loss of IHF. However, one of these promoters, fliK-lacZ, exhibited a wild-type cell cycle-regulated pattern of expression in the absence of IHF. Thus, IHF is required for maximal transcription of several late flagellar genes. The DeltaihfB strain was found to express significantly reduced amounts of the class IV flagellin, FljL, as a consequence of reduced transcriptional activity. Our results indicate that the motility defect exhibited by the DeltaihfB strain is most likely attributable to its failure to accumulate the class IV-encoded 27-kDa flagellin subunit, FljL.
在新月柄杆菌的前分裂细胞中,编码鞭毛胞外组分的III类和IV类鞭毛基因在新生的游动细胞区室中进行转录。这种不对称的表达模式归因于依赖σ54的转录激活因子FlbD的区室化活性。此外,这些在特定时间转录的鞭毛启动子在上游FlbD结合位点和启动子序列之间拥有一段DNA结合蛋白整合宿主因子(IHF)的共有序列。在此,我们删除了新月柄杆菌中编码IHFβ亚基的基因ihfB(himD),并研究了其对鞭毛基因表达的影响。ΔihfB菌株在细胞形态上表现出轻微缺陷,运动能力受损。利用鞭毛启动子报告基因融合体,我们观察到III类鞭毛启动子的一个子集的表达水平因IHF缺失而降低。然而,其中一个启动子fliK-lacZ在无IHF时呈现出野生型细胞周期调控的表达模式。因此,IHF是几个晚期鞭毛基因最大转录所必需的。由于转录活性降低,发现ΔihfB菌株表达的IV类鞭毛蛋白FljL的量显著减少。我们的结果表明,ΔihfB菌株表现出的运动缺陷很可能归因于其未能积累IV类编码的27 kDa鞭毛蛋白亚基FljL。