Molecular analysis of the export-switching apparatus of the anti-sigma factor which regulates the bacterial flagellar regulon

调节细菌鞭毛调节子的抗西格玛因子的输出转换装置的分子分析

• 批准号: 11440221
• 负责人: KUTSUKAKE Kazuhiro
• 金额: $ 9.22万
• 依托单位: OKAYAMA UNIVERSITY (2000-2001)Hiroshima University (1999)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11440221/
• 关键词: Bacterial flagella; Transcriptional control; Peptidoglycan; Flagellar hook; Posttranscriptional control; Protein export; Export switch; Anti-sigma factor; 殺菌鞭毛; シグマ因子; 病原性因子; 形態形成; 突然変異体; 分子集合; ムラミダーゼ; ペリプラズム; 蛋白質間相互作用; シャペロン

1. FlgJ protein was shown to have a muramidase activity involved in penetration process of the peptidoglycan layer by the rod structure. FlgA was shown to be a periplasmic chaperone essential for P-ring formation.2. N-terminal one third of the FlgD protein was shown to be involved in modulation of hook assembly, whereas its C terminal portion was found to enhance the translation and export of the hook protein. This suggests that the monitor system of hook completion may depend upon the FlgD-mediated coupling of translation of the hook protein to its export.3. The FlgM export gate was found to be turned on at the initiation step of hook assembly. This suggests that hook assembly may be carried out with the pool of hook protein which has been already incorporated into the export apparatus by the initiation step of hook assembly.4. Northern blotting analysis found a 200-base RNA molecule derived from the flgB mRNA. The length of this RNA corresponds to axial length of the basal body-hook structure, suggesting that this RNA may be involved in the hook length control. Expression analysis of the fliC-lacZ translational fusion revealed that its expression is slightly repressed after hook assembly. This suggests that translation-controlling machinery of the fliC gene may be established on hook completion.

1. FlgJ蛋白被证明具有参与通过棒状结构穿透肽聚糖层的酶活性。FlgA被证明是p环形成所必需的质周伴侣。FlgD蛋白的三分之一的n端被证明参与钩子组装的调节，而其C端部分被发现促进钩子蛋白的翻译和输出。这表明，钩蛋白完成的监测系统可能依赖于flgd介导的钩蛋白翻译到其输出的偶联。发现FlgM出口门在钩装配的起始步骤被打开。这表明，钩组装可能是用钩蛋白池进行的，钩蛋白池已经通过钩组装的起始步骤纳入了出口装置。Northern blotting分析发现一个来自flgB mRNA的200碱基RNA分子。该RNA的长度与基底体钩结构的轴向长度相对应，提示该RNA可能参与钩长度的控制。fli - lacz翻译融合基因的表达分析表明，钩组装后fli - lacz的表达受到轻微抑制。这表明fliC基因的翻译调控机制可能是在钩子完成过程中建立的。

项目成果

Kutsukake, K.: "Bacterial flagellum : A paradigm for biogenesis of transenvelope supramolecular structures"Recent Research Developments in Microbiology. 4. 607-615 (2000)

Kutsukake, K.：“细菌鞭毛：跨包膜超分子结构生物发生的范例”微生物学的最新研究进展。

Shimamoto,T.: "A cryptic melibiose transporter gene possessing a frameshift from Citrobacter freundii"Journal of Biochemistry. 128(in press). (2001)

Shimamoto,T.：“一种神秘的蜜二糖转运蛋白基因，具有来自弗氏柠檬酸杆菌的移码”生物化学杂志。

Kutsukake, K.: "Two novel regulatory genes, fliTand fliZ, in the flagellar regulon of Salmonella"Genes & Genetic Systems. 74(6)(印刷中). (1999)

Kutsukake, K.：“沙门氏菌鞭毛调节子中的两个新调控基因，fliT 和 fliZ”，《基因与遗传系统》74(6)（出版中）。

Tomoyasu, T.: "The ClpXP ATP-dependent protease regulates flagellum synthesis in Salmonella enterica serovar Typhimurium"Journal of Bacteriology. 184. 645-653 (2002)

Tomoyasu, T.：“ClpXP ATP 依赖性蛋白酶调节鼠伤寒沙门氏菌鞭毛合成”细菌学杂志。

Iyoda,S.: "A flagellar gene fliZ regulates the expression of invasion genes and virulence phenotype in Salmonella enterica serovar Typhimurium"Microbial Pathogenesis. 30(2). 81-90 (2001)

Iyoda,S.：“鞭毛基因 fliZ 调节鼠伤寒沙门氏菌入侵基因的表达和毒力表型”微生物发病机制。