Chen L, Helmann J D
Section of Microbiology, Cornell University, Ithaca, New York 14853.
J Bacteriol. 1994 Jun;176(11):3093-101. doi: 10.1128/jb.176.11.3093-3101.1994.
During a genetic screen to identify metalloregulated loci in Bacillus subtilis, we isolated a Tn917-lacZ insertion in the second gene of an operon downstream of the flagellin (hag) gene. Sequence analysis indicates that this gene encodes a homolog of the enteric flagellar filament cap protein FliD. The fliD gene is followed by homologs of the fliS and fliT genes. Transcription of the fliD-lacZ fusion is sigma D dependent, with peak expression at the end of logarithmic-phase growth. Like other sigma D-dependent genes, expression of fliD-lacZ is greatly reduced by mutations in genes essential for assembly and function of the basal body and hook complex (class II functions). These results suggest that B. subtilis flagellar genes are organized in a hierarchy of gene expression similar to that found in enteric bacteria with hag and fliD as class III genes. Expression from the fliD operon promoter, but not the hag promoter, is repressed by iron, which suggests that the target of metalloregulation is the promoter rather than the sigma D protein.
在一项旨在鉴定枯草芽孢杆菌中金属调节基因座的遗传筛选过程中,我们在鞭毛蛋白(hag)基因下游一个操纵子的第二个基因中分离到一个Tn917 - lacZ插入突变。序列分析表明,该基因编码肠道细菌鞭毛丝帽蛋白FliD的同源物。fliD基因之后是fliS和fliT基因的同源物。fliD - lacZ融合基因的转录依赖于σD,在对数生长期结束时表达达到峰值。与其他依赖σD的基因一样,fliD - lacZ的表达在基体和钩形复合体组装及功能所必需的基因发生突变时(II类功能)会大幅降低。这些结果表明,枯草芽孢杆菌的鞭毛基因按照类似于肠道细菌的基因表达层次进行组织,其中hag和fliD作为III类基因。铁会抑制fliD操纵子启动子的表达,但不影响hag启动子的表达,这表明金属调节的靶点是启动子而非σD蛋白。