Intrinsic clustering of flagellar basal body proteins in E. coli: A self-organization mechanism for assembly and regulation

The assembly and spatial organization of flagellar basal bodies in Escherichia coli are crucial for motility and chemotaxis. Using fluorescence and single-molecule microscopy, we demonstrate that key basal body proteins, FliF and FlhA, self-organize into clusters from low to high expression conditio...

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Main Authors: Yun-Sing Sung, De-Fa Hong, Yi-Ren Chang
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Biochemistry and Biophysics Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2405580825001384
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author Yun-Sing Sung
De-Fa Hong
Yi-Ren Chang
author_facet Yun-Sing Sung
De-Fa Hong
Yi-Ren Chang
author_sort Yun-Sing Sung
collection DOAJ
description The assembly and spatial organization of flagellar basal bodies in Escherichia coli are crucial for motility and chemotaxis. Using fluorescence and single-molecule microscopy, we demonstrate that key basal body proteins, FliF and FlhA, self-organize into clusters from low to high expression conditions. Rather than forming new basal bodies, excess proteins accumulate around pre-existing structures, suggesting an autocatalytic mechanism. It is confirmed that clustering occurs even at low protein levels, indicating an intrinsic organizational principle rather than an artifact of overexpression. Fluorescence recovery after photobleaching (FRAP) revealed dynamic protein exchange within clusters, supporting a diffusion-capture model. Single-molecule analysis showed that FlhA actively remodels clusters, while FliF stabilizes them. 3D imaging suggested that basal body positioning optimizes flagellar distribution for efficient motility. These findings highlight a robust mechanism that regulates basal body positioning and flagellar assembly, ensuring adaptability to varying cellular conditions.
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spelling doaj-art-e969be7225fc4f2a84f10669fd9058172025-08-20T02:37:46ZengElsevierBiochemistry and Biophysics Reports2405-58082025-06-014210205110.1016/j.bbrep.2025.102051Intrinsic clustering of flagellar basal body proteins in E. coli: A self-organization mechanism for assembly and regulationYun-Sing Sung0De-Fa Hong1Yi-Ren Chang2Department of Physics, National Taiwan Normal University, Taipei, 116, TaiwanDepartment of Physics, National Taiwan Normal University, Taipei, 116, TaiwanCorresponding author. 88, Sec.4, Ting-Chou Rd., Taipei, 116, Taiwan.; Department of Physics, National Taiwan Normal University, Taipei, 116, TaiwanThe assembly and spatial organization of flagellar basal bodies in Escherichia coli are crucial for motility and chemotaxis. Using fluorescence and single-molecule microscopy, we demonstrate that key basal body proteins, FliF and FlhA, self-organize into clusters from low to high expression conditions. Rather than forming new basal bodies, excess proteins accumulate around pre-existing structures, suggesting an autocatalytic mechanism. It is confirmed that clustering occurs even at low protein levels, indicating an intrinsic organizational principle rather than an artifact of overexpression. Fluorescence recovery after photobleaching (FRAP) revealed dynamic protein exchange within clusters, supporting a diffusion-capture model. Single-molecule analysis showed that FlhA actively remodels clusters, while FliF stabilizes them. 3D imaging suggested that basal body positioning optimizes flagellar distribution for efficient motility. These findings highlight a robust mechanism that regulates basal body positioning and flagellar assembly, ensuring adaptability to varying cellular conditions.http://www.sciencedirect.com/science/article/pii/S2405580825001384Flagellar assemblyProtein self-organizationSpatial regulationFluorescence super-resolution imagingSingle-molecule methods
spellingShingle Yun-Sing Sung
De-Fa Hong
Yi-Ren Chang
Intrinsic clustering of flagellar basal body proteins in E. coli: A self-organization mechanism for assembly and regulation
Biochemistry and Biophysics Reports
Flagellar assembly
Protein self-organization
Spatial regulation
Fluorescence super-resolution imaging
Single-molecule methods
title Intrinsic clustering of flagellar basal body proteins in E. coli: A self-organization mechanism for assembly and regulation
title_full Intrinsic clustering of flagellar basal body proteins in E. coli: A self-organization mechanism for assembly and regulation
title_fullStr Intrinsic clustering of flagellar basal body proteins in E. coli: A self-organization mechanism for assembly and regulation
title_full_unstemmed Intrinsic clustering of flagellar basal body proteins in E. coli: A self-organization mechanism for assembly and regulation
title_short Intrinsic clustering of flagellar basal body proteins in E. coli: A self-organization mechanism for assembly and regulation
title_sort intrinsic clustering of flagellar basal body proteins in e coli a self organization mechanism for assembly and regulation
topic Flagellar assembly
Protein self-organization
Spatial regulation
Fluorescence super-resolution imaging
Single-molecule methods
url http://www.sciencedirect.com/science/article/pii/S2405580825001384
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