In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles
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-regula...
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MDPI AG
2020-01-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/1/126 |
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| author | Hiroyuki Terashima Chinatsu Tatsumi Akihiro Kawamoto Keiichi Namba Tohru Minamino Katsumi Imada |
| author_facet | Hiroyuki Terashima Chinatsu Tatsumi Akihiro Kawamoto Keiichi Namba Tohru Minamino Katsumi Imada |
| author_sort | Hiroyuki Terashima |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-19T22:02:31Z |
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| id | doaj.art-39081fa3def34f6fb5146ee72653b990 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-12-19T22:02:31Z |
| publishDate | 2020-01-01 |
| publisher | MDPI AG |
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| series | Biomolecules |
| spelling | doaj.art-39081fa3def34f6fb5146ee72653b9902022-12-21T20:04:06ZengMDPI AGBiomolecules2218-273X2020-01-0110112610.3390/biom10010126biom10010126In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane VesiclesHiroyuki Terashima0Chinatsu Tatsumi1Akihiro Kawamoto2Keiichi Namba3Tohru Minamino4Katsumi Imada5Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-Ku, Nagoya 464-8602, JapanDepartment of Macromolecular Science, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanGraduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, JapanGraduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, JapanGraduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, JapanDepartment of Macromolecular Science, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanThe 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.https://www.mdpi.com/2218-273X/10/1/126bacterial flagellumtype iii secretion systemflagellar specific export apparatusinverted membrane vesiclein vitro reconstitutionflagellar filament<i>salmonella typhimurium</i> |
| spellingShingle | Hiroyuki Terashima Chinatsu Tatsumi Akihiro Kawamoto Keiichi Namba Tohru Minamino Katsumi Imada In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles Biomolecules bacterial flagellum type iii secretion system flagellar specific export apparatus inverted membrane vesicle in vitro reconstitution flagellar filament <i>salmonella typhimurium</i> |
| title | In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles |
| title_full | In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles |
| title_fullStr | In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles |
| title_full_unstemmed | In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles |
| title_short | In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles |
| title_sort | in vitro autonomous construction of the flagellar axial structure in inverted membrane vesicles |
| topic | bacterial flagellum type iii secretion system flagellar specific export apparatus inverted membrane vesicle in vitro reconstitution flagellar filament <i>salmonella typhimurium</i> |
| url | https://www.mdpi.com/2218-273X/10/1/126 |
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