Kutsukake K, Okada T, Yokoseki T, Iino T
Faculty of Applied Biological Science, Hiroshima University, Japan.
Gene. 1994 May 27;143(1):49-54. doi: 10.1016/0378-1119(94)90603-3.
The flagellar genes flgA and flgM are located at the terminus of the region-I flagellar gene cluster on the chromosome of Salmonella typhimurium. The flgA gene is involved in P-ring formation of the flagellar basal body, whereas flgM encodes the anti-sigma factor which acts as a negative regulator of the flagellar regulon. The nucleotide sequence of the DNA fragment containing these flagellar genes and the adjacent region was determined. The flgA gene was found to encode a 219-amino-acid (aa) protein of 23,556 Da. The N-terminal region of FlgA has the characteristics of a typical signal sequence, suggesting that FlgA may function in the periplasmic space where P-ring assembly takes place. The flgM gene was found to constitute an operon together with an ORF which encodes a 140-aa protein of 15,899 Da. A gene disruption mutant was constructed by inserting a cat gene cartridge into the ORF on the chromosome. This mutant showed only weak motility, indicating that the product of the ORF is involved in flagellar formation. Therefore, this ORF was designated as flgN. Electron microscopic observation revealed that most of the flagellar structures produced by the flgN mutant are hook-basal body complexes lacking the filament portions. Based on these results, we concluded that the flgN product is required for the efficient initiation of filament assembly.
鞭毛基因flgA和flgM位于鼠伤寒沙门氏菌染色体上区域I鞭毛基因簇的末端。flgA基因参与鞭毛基体P环的形成,而flgM编码抗σ因子,该因子作为鞭毛调节子的负调控因子。测定了包含这些鞭毛基因及相邻区域的DNA片段的核苷酸序列。发现flgA基因编码一个由219个氨基酸组成、分子量为23556 Da的蛋白质。FlgA的N端区域具有典型信号序列的特征,表明FlgA可能在P环组装发生的周质空间发挥作用。发现flgM基因与一个编码140个氨基酸、分子量为15899 Da的蛋白质的开放阅读框(ORF)共同构成一个操纵子。通过将一个氯霉素基因盒插入染色体上的ORF构建了一个基因破坏突变体。该突变体仅表现出微弱的运动性,表明该ORF的产物参与鞭毛形成。因此,将这个ORF命名为flgN。电子显微镜观察显示,flgN突变体产生的大多数鞭毛结构是缺少丝状体部分的钩-基体复合体。基于这些结果,我们得出结论,flgN产物是丝状体组装有效起始所必需的。