A Conserved Cell-pole Determinant Organizes Proper Polar Flagellum Formation

Overview

Journal Elife

Specialty Biology

Date 2024 Dec 5

PMID 39636223

Authors

Erick E Arroyo-Perez

John C Hook

Alejandra Alvarado

Stephan Wimmi

Timo Glatter

Kai Thormann

Simon Ringgaard

Affiliations

Soon will be listed here.

Abstract

The coordination of cell cycle progression and flagellar synthesis is a complex process in motile bacteria. In γ-proteobacteria, the localization of the flagellum to the cell pole is mediated by the SRP-type GTPase FlhF. However, the mechanism of action of FlhF, and its relationship with the cell pole landmark protein HubP remain unclear. In this study, we discovered a novel protein called FipA that is required for normal FlhF activity and function in polar flagellar synthesis. We demonstrated that membrane-localized FipA interacts with FlhF and is required for normal flagellar synthesis in , , and , and it does so independently of the polar localization mediated by HubP. FipA exhibits a dynamic localization pattern and is present at the designated pole before flagellar synthesis begins, suggesting its role in licensing flagellar formation. This discovery provides insight into a new pathway for regulating flagellum synthesis and coordinating cellular organization in bacteria that rely on polar flagellation and FlhF-dependent localization.

References

Muraleedharan S, Freitas C, Mann P, Glatter T, Ringgaard S . A cell length-dependent transition in MinD-dynamics promotes a switch in division-site placement and preservation of proliferating elongated Vibrio parahaemolyticus swarmer cells. Mol Microbiol. 2018; 109(3):365-384. DOI: 10.1111/mmi.13996. View

Kusumoto A, Kamisaka K, Yakushi T, Terashima H, Shinohara A, Homma M . Regulation of polar flagellar number by the flhF and flhG genes in Vibrio alginolyticus. J Biochem. 2006; 139(1):113-21. DOI: 10.1093/jb/mvj010. View

Dornes A, Schmidt L, Mais C, Hook J, Pane-Farre J, Kressler D . Polar confinement of a macromolecular machine by an SRP-type GTPase. Nat Commun. 2024; 15(1):5797. PMC: 11236974. DOI: 10.1038/s41467-024-50274-4. View

Chen M, Zhao Z, Yang J, Peng K, Baker M, Bai F . Length-dependent flagellar growth of revealed by real time fluorescent imaging. Elife. 2017; 6. PMC: 5300704. DOI: 10.7554/eLife.22140. View

Coil D, Anne J . The role of fimV and the importance of its tandem repeat copy number in twitching motility, pigment production, and morphology in Legionella pneumophila. Arch Microbiol. 2010; 192(8):625-31. DOI: 10.1007/s00203-010-0590-8. View

Hendrixson D, DiRita V . Transcription of sigma54-dependent but not sigma28-dependent flagellar genes in Campylobacter jejuni is associated with formation of the flagellar secretory apparatus. Mol Microbiol. 2003; 50(2):687-702. DOI: 10.1046/j.1365-2958.2003.03731.x. View

Gibson D, Young L, Chuang R, Venter J, Hutchison 3rd C, Smith H . Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods. 2009; 6(5):343-5. DOI: 10.1038/nmeth.1318. View

Hook J, Blagotinsek V, Pane-Farre J, Mrusek D, Altegoer F, Dornes A . A Proline-Rich Element in the Type III Secretion Protein FlhB Contributes to Flagellar Biogenesis in the Beta- and Gamma-Proteobacteria. Front Microbiol. 2021; 11:564161. PMC: 7771051. DOI: 10.3389/fmicb.2020.564161. View

Nelson K, Weinel C, Paulsen I, Dodson R, HILBERT H, Martins Dos Santos V . Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440. Environ Microbiol. 2003; 4(12):799-808. DOI: 10.1046/j.1462-2920.2002.00366.x. View

10.

Blagotinsek V, Schwan M, Steinchen W, Mrusek D, Hook J, Rossmann F . An ATP-dependent partner switch links flagellar C-ring assembly with gene expression. Proc Natl Acad Sci U S A. 2020; 117(34):20826-20835. PMC: 7456076. DOI: 10.1073/pnas.2006470117. View

11.

Green J, Kahramanoglou C, Rahman A, Pender A, Charbonnel N, Fraser G . Recruitment of the earliest component of the bacterial flagellum to the old cell division pole by a membrane-associated signal recognition particle family GTP-binding protein. J Mol Biol. 2009; 391(4):679-90. DOI: 10.1016/j.jmb.2009.05.075. View

12.

Campos-Garcia J, Najera R, Camarena L, Soberon-Chavez G . The pseudomonas aeruginosa motR gene involved in regulation of bacterial motility. FEMS Microbiol Lett. 2000; 184(1):57-62. DOI: 10.1111/j.1574-6968.2000.tb08990.x. View

13.

Kim Y, McCarter L . Analysis of the polar flagellar gene system of Vibrio parahaemolyticus. J Bacteriol. 2000; 182(13):3693-704. PMC: 94540. DOI: 10.1128/JB.182.13.3693-3704.2000. View

14.

Schuhmacher J, Rossmann F, Dempwolff F, Knauer C, Altegoer F, Steinchen W . MinD-like ATPase FlhG effects location and number of bacterial flagella during C-ring assembly. Proc Natl Acad Sci U S A. 2015; 112(10):3092-7. PMC: 4364217. DOI: 10.1073/pnas.1419388112. View

15.

Kusumoto A, Nishioka N, Kojima S, Homma M . Mutational analysis of the GTP-binding motif of FlhF which regulates the number and placement of the polar flagellum in Vibrio alginolyticus. J Biochem. 2009; 146(5):643-50. DOI: 10.1093/jb/mvp109. View

16.

Rossmann F, Brenzinger S, Knauer C, Dorrich A, Bubendorfer S, Ruppert U . The role of FlhF and HubP as polar landmark proteins in Shewanella putrefaciens CN-32. Mol Microbiol. 2015; 98(4):727-42. DOI: 10.1111/mmi.13152. View

17.

Francis N, Sosinsky G, Thomas D, DeRosier D . Isolation, characterization and structure of bacterial flagellar motors containing the switch complex. J Mol Biol. 1994; 235(4):1261-70. DOI: 10.1006/jmbi.1994.1079. View

18.

Macnab R . How bacteria assemble flagella. Annu Rev Microbiol. 2003; 57:77-100. DOI: 10.1146/annurev.micro.57.030502.090832. View

19.

Dasgupta N, Ramphal R . Interaction of the antiactivator FleN with the transcriptional activator FleQ regulates flagellar number in Pseudomonas aeruginosa. J Bacteriol. 2001; 183(22):6636-44. PMC: 95495. DOI: 10.1128/JB.183.22.6636-6644.2001. View

20.

Karimova G, Pidoux J, Ullmann A, Ladant D . A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc Natl Acad Sci U S A. 1998; 95(10):5752-6. PMC: 20451. DOI: 10.1073/pnas.95.10.5752. View