Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion
Flagella-driven motility enables bacteria to reach their favorable niche within the host. The human foodborne pathogen Campylobacter jejuni produces two heavily glycosylated structural flagellins (FlaA and FlaB) that form the flagellar filament. It also encodes the non-structural FlaC flagellin whic...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-06-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2017.01060/full |
| _version_ | 1811204763866365952 |
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| author | Katarzyna A. Radomska Marc M. S. M. Wösten Soledad R. Ordoñez Jaap A. Wagenaar Jaap A. Wagenaar Jaap A. Wagenaar Jos P. M. van Putten Jos P. M. van Putten |
| author_facet | Katarzyna A. Radomska Marc M. S. M. Wösten Soledad R. Ordoñez Jaap A. Wagenaar Jaap A. Wagenaar Jaap A. Wagenaar Jos P. M. van Putten Jos P. M. van Putten |
| author_sort | Katarzyna A. Radomska |
| collection | DOAJ |
| description | Flagella-driven motility enables bacteria to reach their favorable niche within the host. The human foodborne pathogen Campylobacter jejuni produces two heavily glycosylated structural flagellins (FlaA and FlaB) that form the flagellar filament. It also encodes the non-structural FlaC flagellin which is secreted through the flagellum and has been implicated in host cell invasion. The mechanisms that regulate C. jejuni flagellin biogenesis and guide the proteins to the export apparatus are different from those in most other enteropathogens and are not fully understood. This work demonstrates the importance of the putative flagellar protein FliS in C. jejuni flagella assembly. A constructed fliS knockout strain was non-motile, displayed reduced levels of FlaA/B and FlaC flagellin, and carried severely truncated flagella. Pull-down and Far Western blot assays showed direct interaction of FliS with all three C. jejuni flagellins (FlaA, FlaB, and FlaC). This is in contrast to, the sensor and regulator of intracellular flagellin levels, FliW, which bound to FlaA and FlaB but not to FlaC. The FliS protein but not FliW preferred binding to glycosylated C. jejuni flagellins rather than to their non-glycosylated recombinant counterparts. Mapping of the binding region of FliS and FliW using a set of flagellin fragments showed that the C-terminal subdomain of the flagellin was required for FliS binding, whereas the N-terminal subdomain was essential for FliW binding. The separate binding subdomains required for FliS and FliW, the different substrate specificity, and the differential preference for binding of glycosylated flagellins ensure optimal processing and assembly of the C. jejuni flagellins. |
| first_indexed | 2024-04-12T03:19:14Z |
| format | Article |
| id | doaj.art-002b632999bf4c23a74ebeee83f090a9 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-12T03:19:14Z |
| publishDate | 2017-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-002b632999bf4c23a74ebeee83f090a92022-12-22T03:49:57ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2017-06-01810.3389/fmicb.2017.01060262965Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin SecretionKatarzyna A. Radomska0Marc M. S. M. Wösten1Soledad R. Ordoñez2Jaap A. Wagenaar3Jaap A. Wagenaar4Jaap A. Wagenaar5Jos P. M. van Putten6Jos P. M. van Putten7Department of Infectious Diseases and Immunology, Utrecht UniversityUtrecht, NetherlandsDepartment of Infectious Diseases and Immunology, Utrecht UniversityUtrecht, NetherlandsDepartment of Infectious Diseases and Immunology, Utrecht UniversityUtrecht, NetherlandsDepartment of Infectious Diseases and Immunology, Utrecht UniversityUtrecht, NetherlandsWageningen Bioveterinary ResearchLelystad, NetherlandsWHO Collaborating Centre for Campylobacter/OIE Reference Laboratory for CampylobacteriosisUtrecht, NetherlandsDepartment of Infectious Diseases and Immunology, Utrecht UniversityUtrecht, NetherlandsWHO Collaborating Centre for Campylobacter/OIE Reference Laboratory for CampylobacteriosisUtrecht, NetherlandsFlagella-driven motility enables bacteria to reach their favorable niche within the host. The human foodborne pathogen Campylobacter jejuni produces two heavily glycosylated structural flagellins (FlaA and FlaB) that form the flagellar filament. It also encodes the non-structural FlaC flagellin which is secreted through the flagellum and has been implicated in host cell invasion. The mechanisms that regulate C. jejuni flagellin biogenesis and guide the proteins to the export apparatus are different from those in most other enteropathogens and are not fully understood. This work demonstrates the importance of the putative flagellar protein FliS in C. jejuni flagella assembly. A constructed fliS knockout strain was non-motile, displayed reduced levels of FlaA/B and FlaC flagellin, and carried severely truncated flagella. Pull-down and Far Western blot assays showed direct interaction of FliS with all three C. jejuni flagellins (FlaA, FlaB, and FlaC). This is in contrast to, the sensor and regulator of intracellular flagellin levels, FliW, which bound to FlaA and FlaB but not to FlaC. The FliS protein but not FliW preferred binding to glycosylated C. jejuni flagellins rather than to their non-glycosylated recombinant counterparts. Mapping of the binding region of FliS and FliW using a set of flagellin fragments showed that the C-terminal subdomain of the flagellin was required for FliS binding, whereas the N-terminal subdomain was essential for FliW binding. The separate binding subdomains required for FliS and FliW, the different substrate specificity, and the differential preference for binding of glycosylated flagellins ensure optimal processing and assembly of the C. jejuni flagellins.http://journal.frontiersin.org/article/10.3389/fmicb.2017.01060/fullCampylobacter jejuniflagellar motilityflagellar chaperoneflagellinFliSFliW |
| spellingShingle | Katarzyna A. Radomska Marc M. S. M. Wösten Soledad R. Ordoñez Jaap A. Wagenaar Jaap A. Wagenaar Jaap A. Wagenaar Jos P. M. van Putten Jos P. M. van Putten Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion Frontiers in Microbiology Campylobacter jejuni flagellar motility flagellar chaperone flagellin FliS FliW |
| title | Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion |
| title_full | Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion |
| title_fullStr | Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion |
| title_full_unstemmed | Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion |
| title_short | Importance of Campylobacter jejuni FliS and FliW in Flagella Biogenesis and Flagellin Secretion |
| title_sort | importance of campylobacter jejuni flis and fliw in flagella biogenesis and flagellin secretion |
| topic | Campylobacter jejuni flagellar motility flagellar chaperone flagellin FliS FliW |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2017.01060/full |
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