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Ring formation by Vibrio fusion protein composed of FliF and FliG, MS-ring and C-ring component of bacterial flagellar motor in membrane. 由 FliF 和 FliG 组成的弧菌融合蛋白、MS 环和细菌鞭毛运动的 C 环在膜中形成环。

Biophysics and physicobiology Pub Date : 2023-06-09 eCollection Date: 2023-01-01 DOI: 10.2142/biophysico.bppb-v20.0028

Kanji Takahashi, Tatsuro Nishikino, Hiroki Kajino, Seiji Kojima, Takayuki Uchihashi, Michio Homma

{"title":"Ring formation by Vibrio fusion protein composed of FliF and FliG, MS-ring and C-ring component of bacterial flagellar motor in membrane.","authors":"Kanji Takahashi, Tatsuro Nishikino, Hiroki Kajino, Seiji Kojima, Takayuki Uchihashi, Michio Homma","doi":"10.2142/biophysico.bppb-v20.0028","DOIUrl":"10.2142/biophysico.bppb-v20.0028","url":null,"abstract":"The marine bacterium Vibrio alginolyticus has a single flagellum as a locomotory organ at the cell pole, which is rotated by the Na+-motive force to swim in a liquid. The base of the flagella has a motor composed of a stator and rotor, which serves as a power engine to generate torque through the rotor-stator interaction coupled to Na+ influx through the stator channel. The MS-ring, which is embedded in the membrane at the base of the flagella as part of the rotor, is the initial structure required for flagellum assembly. It comprises 34 molecules of the two-transmembrane protein FliF. FliG, FliM, and FliN form a C-ring just below the MS-ring. FliG is an important rotor protein that interacts with the stator PomA and directly contributes to force generation. We previously found that FliG promotes MS-ring formation in E. coli. In the present study, we constructed a fliF-fliG fusion gene, which encodes an approximately 100 kDa protein, and the successful production of this protein effectively formed the MS-ring in E. coli cells. We observed fuzzy structures around the ring using either electron microscopy or high-speed atomic force microscopy (HS-AFM), suggesting that FliM and FliN are necessary for the formation of a stable ring structure. The HS-AFM movies revealed flexible movements at the FliG region.","PeriodicalId":101323,"journal":{"name":"Biophysics and physicobiology","volume":"20 2","pages":"e200028"},"PeriodicalIF":0.0,"publicationDate":"2023-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10941966/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140145021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

RNA interference reveals the escape response mechanism of Paramecium to mechanical stimulation. RNA干扰揭示草履虫对机械刺激的逃逸反应机制。

Biophysics and physicobiology Pub Date : 2023-05-31 eCollection Date: 2023-06-14 DOI: 10.2142/biophysico.bppb-v20.0025

Manabu Hori, Takashi Tominaga, Masaki Ishida, Mutsumi Kawano

{"title":"RNA interference reveals the escape response mechanism of Paramecium to mechanical stimulation.","authors":"Manabu Hori, Takashi Tominaga, Masaki Ishida, Mutsumi Kawano","doi":"10.2142/biophysico.bppb-v20.0025","DOIUrl":"10.2142/biophysico.bppb-v20.0025","url":null,"abstract":"In Paramecium, a mechanical stimulus applied to the posterior portion of the cell causes a transient increase in membrane permeability to potassium ions, transiently rendering the membrane in a hyperpolarized state. Hyperpolarization causes a transient increase in Cyclic adenosine monophosphate (cAMP) concentration in the cilia, resulting in a transient fast-forward swimming of the cell. Schultz and coworkers (1992) reported that a unique adenylate cyclase (AC)-coupled potassium channel is involved in the reaction underlying this response, which is known as the \"escape response.\" However, the AC responsible for this reaction remains to be identified. Moreover, the molecular linkage between mechanoreception and AC activation has not been elucidated adequately. Currently, we can perform an efficient and simple gene-knockdown technique in Paramecium using RNA interference (RNAi). Paramecium is one of the several model organisms for which whole-genome sequences have been elucidated. The RNAi technique can be applied to whole genome sequences derived from the Paramecium database (ParameciumDB) to investigate the types of proteins that elicit specific biological responses and compare them with those of other model organisms. In this review, we describe the applications of the RNAi technique in elucidating the molecular mechanism underlying the escape response and identifying the AC involved in this reaction. The findings of this study highlight the advantages of the RNAi technique and ParameciumDB.","PeriodicalId":101323,"journal":{"name":"Biophysics and physicobiology","volume":"20 2","pages":"e200025"},"PeriodicalIF":0.0,"publicationDate":"2023-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ca/89/20_e200025.PMC10587447.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49695849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Flagellar polymorphism-dependent bacterial swimming motility in a structured environment. 结构环境中鞭毛多态性依赖的细菌游泳运动性。

Biophysics and physicobiology Pub Date : 2023-05-30 eCollection Date: 2023-06-14 DOI: 10.2142/biophysico.bppb-v20.0024

Yoshiaki Kinosita, Yoshiyuki Sowa

{"title":"Flagellar polymorphism-dependent bacterial swimming motility in a structured environment.","authors":"Yoshiaki Kinosita, Yoshiyuki Sowa","doi":"10.2142/biophysico.bppb-v20.0024","DOIUrl":"10.2142/biophysico.bppb-v20.0024","url":null,"abstract":"Most motile bacteria use supramolecular motility machinery called bacterial flagellum, which converts the chemical energy gained from ion flux into mechanical rotation. Bacterial cells sense their external environment through a two-component regulatory system consisting of a histidine kinase and response regulator. Combining these systems allows the cells to move toward favorable environments and away from their repellents. A representative example of flagellar motility is run-and-tumble swimming in Escherichia coli, where the counter-clockwise (CCW) rotation of a flagellar bundle propels the cell forward, and the clockwise (CW) rotation undergoes cell re-orientation (tumbling) upon switching the direction of flagellar motor rotation from CCW to CW. In this mini review, we focus on several types of chemotactic behaviors that respond to changes in flagellar shape and direction of rotation. Moreover, our single-cell analysis demonstrated back-and-forth swimming motility of an original E. coli strain. We propose that polymorphic flagellar changes are required to enhance bacterial movement in a structured environment as a colony spread on an agar plate.","PeriodicalId":101323,"journal":{"name":"Biophysics and physicobiology","volume":"20 2","pages":"e200024"},"PeriodicalIF":0.0,"publicationDate":"2023-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/32/e2/20_e200024.PMC10587448.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49695848","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

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