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蛋白具有胞壁质酶活性，参与杆状结构对肽聚糖层的渗透过程。 FlgA被证明是P环形成所必需的周质伴侣。2． FlgD 蛋白的 N 端三分之一参与了钩子组装的调节，而其 C 端部分被发现可增强钩子蛋白的翻译和输出。这表明钩完成的监测系统可能依赖于FlgD介导的钩蛋白翻译与其输出的耦合。3．发现 FlgM 出口门在钩组装的启动步骤中打开。这表明可以用已经通过钩组装的起始步骤并入输出装置中的钩蛋白质池来进行钩组装。4． Northern 印迹分析发现源自 flgB mRNA 的 200 个碱基的 RNA 分子。该RNA的长度对应于基底体-钩结构的轴向长度，表明该RNA可能参与钩长度控制。 fliC-lacZ 翻译融合体的表达分析表明，其表达在钩子组装后受到轻微抑制。这表明 fliC 基因的翻译控制机制可能是在钩子完成时建立的。

项目成果

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。

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）。

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。