Faculty Collaboration Database

Borrelia burgdorferi tolerates alteration to P66 porin function in a murine infectivity model. Front Cell Infect Microbiol 2024;14:1528456

Date

02/05/2025

Pubmed ID

39906208

Pubmed Central ID

PMC11790652

DOI

10.3389/fcimb.2024.1528456

Scopus ID

2-s2.0-85216789061 (requires institutional sign-in at Scopus site)   1 Citation

Abstract

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.

Author List

Fierros CH, Faucillion ML, Hahn BL, Anderson P, Bonde M, Kessler JR, Surdel MC, Crawford KS, Gao Y, Zhu J, Bergström S, Coburn J

Authors

Jenifer Coburn PhD, MA Professor in the Medicine department at Medical College of Wisconsin
Yan Gao PhD Assistant Professor in the Data Science Institute department at Medical College of Wisconsin
Matthew C. Surdel PhD Assistant Professor in the Medicine department at Medical College of Wisconsin
Jieqing Zhu PhD Professor in the Biochemistry department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Bacterial Proteins
Borrelia burgdorferi
Disease Models, Animal
Female
Lyme Disease
Mice
Mutagenesis, Site-Directed
Mutation, Missense
Porins
Virulence