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Open AccessArticle

In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles

1
Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-Ku, Nagoya 464-8602, Japan
2
Department of Macromolecular Science, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, Japan
3
Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan
4
RIKEN SPring-8 Center and Center for Biosystems Dynamic Research, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan
5
JEOL YOKOGUSHI Research Alliance Laboratories, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan
*
Author to whom correspondence should be addressed.
Current Affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Biomolecules 2020, 10(1), 126; https://doi.org/10.3390/biom10010126
Received: 16 December 2019 / Revised: 8 January 2020 / Accepted: 9 January 2020 / Published: 11 January 2020
(This article belongs to the Special Issue Perspectives on Bacterial Flagellar Motor)
The bacterial flagellum is a filamentous organelle extending from the cell surface. The axial structure of the flagellum consists of the rod, hook, junction, filament, and cap. The axial structure is formed by axial component proteins exported via a specific protein export apparatus in a well-regulated manner. Although previous studies have revealed the outline of the flagellar construction process, the mechanism of axial structure formation, including axial protein export, is still obscure due to difficulties in direct observation of protein export and assembly in vivo. We recently developed an in vitro flagellar protein transport assay system using inverted membrane vesicles (IMVs) and succeeded in reproducing the early stage of flagellar assembly. However, the late stage of the flagellar formation process remained to be examined in the IMVs. In this study, we showed that the filament-type proteins are transported into the IMVs to produce the filament on the hook inside the IMVs. Furthermore, we provide direct evidence that coordinated flagellar protein export and assembly can occur at the post-translational level. These results indicate that the ordered construction of the entire flagellar structure can be regulated by only the interactions between the protein export apparatus, the export substrate proteins, and their cognate chaperones. View Full-Text
Keywords: bacterial flagellum; type III secretion system; flagellar specific export apparatus; inverted membrane vesicle; in vitro reconstitution; flagellar filament; Salmonella typhimurium bacterial flagellum; type III secretion system; flagellar specific export apparatus; inverted membrane vesicle; in vitro reconstitution; flagellar filament; Salmonella typhimurium
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Terashima, H.; Tatsumi, C.; Kawamoto, A.; Namba, K.; Minamino, T.; Imada, K. In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles. Biomolecules 2020, 10, 126.

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