Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity model
Borrelia burgdorferi exists in a complex enzootic life cycle requiring differential gene regulation. P66, a porin and adhesin, is upregulated and essential during mammalian infection, but is not produced or required within the tick vector. We sought to determine whether the porin function of P66 is...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2024.1528456/full |
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| author | Christa H. Fierros Marie-Line Faucillion Beth L. Hahn Phillip Anderson Mari Bonde Julie R. Kessler Matthew C. Surdel Kyler S. Crawford Yan Gao Jieqing Zhu Jieqing Zhu Sven Bergström Jenifer Coburn Jenifer Coburn |
| author_facet | Christa H. Fierros Marie-Line Faucillion Beth L. Hahn Phillip Anderson Mari Bonde Julie R. Kessler Matthew C. Surdel Kyler S. Crawford Yan Gao Jieqing Zhu Jieqing Zhu Sven Bergström Jenifer Coburn Jenifer Coburn |
| author_sort | Christa H. Fierros |
| collection | DOAJ |
| description | Borrelia burgdorferi exists in a complex enzootic life cycle requiring differential gene regulation. P66, a porin and adhesin, is upregulated and essential during mammalian infection, but is not produced or required within the tick vector. We sought to determine whether the porin function of P66 is essential for infection. Vancomycin treatment of B. burgdorferi cultures was used to screen for P66 porin function and found to generate spontaneous mutations in p66 (bb0603). Three novel, spontaneous, missense P66 mutants (G175V, T176M, and G584R) were re-created by site-directed mutagenesis in an infectious strain background and tested for infectivity in mice by ID50 experiments. Two of the three mutants retained infectivity comparable to the isogenic control, suggesting that B. burgdorferi can tolerate alteration to P66 porin function during infection. The third mutant exhibited highly attenuated infectivity and produced low levels of P66 protein. Interestingly, four isolates that were recovered for p66 sequencing from mouse tissues revealed novel secondary point mutations in genomic p66. However, these secondary mutations did not rescue P66 porin function. New structural modeling of P66 is presented and consistent with these experimental results. This is the first work to assess the contribution of P66 porin function to B. burgdorferi pathogenesis. |
| format | Article |
| id | doaj-art-4b4fbf90d6264269ba23928bd0558d09 |
| institution | Kabale University |
| issn | 2235-2988 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj-art-4b4fbf90d6264269ba23928bd0558d092025-01-21T08:36:32ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-01-011410.3389/fcimb.2024.15284561528456Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity modelChrista H. Fierros0Marie-Line Faucillion1Beth L. Hahn2Phillip Anderson3Mari Bonde4Julie R. Kessler5Matthew C. Surdel6Kyler S. Crawford7Yan Gao8Jieqing Zhu9Jieqing Zhu10Sven Bergström11Jenifer Coburn12Jenifer Coburn13Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Molecular Biology, Umeå University, Umea, SwedenDepartment of Medicine, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Medicine, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Molecular Biology, Umeå University, Umea, SwedenDepartment of Medicine, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Medicine, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, United StatesDivision of Biostatistics, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Biochemistry, Medical College of Wisconsin, Versiti Blood Research Institute, Milwaukee, WI, United StatesDepartment of Molecular Biology, Umeå University, Umea, SwedenDepartment of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Medicine, Medical College of Wisconsin, Milwaukee, WI, United StatesBorrelia burgdorferi exists in a complex enzootic life cycle requiring differential gene regulation. P66, a porin and adhesin, is upregulated and essential during mammalian infection, but is not produced or required within the tick vector. We sought to determine whether the porin function of P66 is essential for infection. Vancomycin treatment of B. burgdorferi cultures was used to screen for P66 porin function and found to generate spontaneous mutations in p66 (bb0603). Three novel, spontaneous, missense P66 mutants (G175V, T176M, and G584R) were re-created by site-directed mutagenesis in an infectious strain background and tested for infectivity in mice by ID50 experiments. Two of the three mutants retained infectivity comparable to the isogenic control, suggesting that B. burgdorferi can tolerate alteration to P66 porin function during infection. The third mutant exhibited highly attenuated infectivity and produced low levels of P66 protein. Interestingly, four isolates that were recovered for p66 sequencing from mouse tissues revealed novel secondary point mutations in genomic p66. However, these secondary mutations did not rescue P66 porin function. New structural modeling of P66 is presented and consistent with these experimental results. This is the first work to assess the contribution of P66 porin function to B. burgdorferi pathogenesis.https://www.frontiersin.org/articles/10.3389/fcimb.2024.1528456/fullP66porinBorrelia burgdorferiLyme diseasebacterial pathogenesistickborne pathogen |
| spellingShingle | Christa H. Fierros Marie-Line Faucillion Beth L. Hahn Phillip Anderson Mari Bonde Julie R. Kessler Matthew C. Surdel Kyler S. Crawford Yan Gao Jieqing Zhu Jieqing Zhu Sven Bergström Jenifer Coburn Jenifer Coburn Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity model Frontiers in Cellular and Infection Microbiology P66 porin Borrelia burgdorferi Lyme disease bacterial pathogenesis tickborne pathogen |
| title | Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity model |
| title_full | Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity model |
| title_fullStr | Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity model |
| title_full_unstemmed | Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity model |
| title_short | Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity model |
| title_sort | borrelia burgdorferi tolerates alteration to p66 porin function in a murine infectivity model |
| topic | P66 porin Borrelia burgdorferi Lyme disease bacterial pathogenesis tickborne pathogen |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2024.1528456/full |
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