Analysis of the cellular localization of Bdr paralogs in Borrelia burgdorferi, a causative agent of lyme disease: evidence for functional diversity.
The bdr (Borrelia direct repeat) gene family of the genus Borrelia encodes a polymorphic group of proteins that carry a central repeat motif region containing putative phosphorylation sites and a hydrophobic carboxyl-terminal domain. It has been postulated that the Bdr proteins may anchor to the inner membrane via the C-terminal domain. In this study, we used cellular fractionation methodologies, salt and detergent treatments, and immunoblot analyses to assess the association of the Bdr proteins with the cellular infrastructure in both Borrelia burgdorferi (a Lyme disease spirochete) and B. turicatae (a relapsing fever spirochete). Triton X-114 extraction and partitioning experiments demonstrated that most Bdr paralogs are associated with the inner membrane-peptidoglycan complex. Analyses of cells treated with the highly chaotropic bile salt detergent deoxycholic acid demonstrated that some Bdr paralogs may also interact with the peptidoglycan, as evidenced by their tight association with the insoluble cellular matrix. In addition, immunoprecipitation (IP) experiments revealed an enhanced IP of all Bdr paralogs when the cell lysates were boiled prior to addition of the precipitating antibody. Furthermore, some Bdr paralogs were accessible to antibody in the IP experiments only in the boiled cell lysates. These observations suggest that different Bdr paralogs may carry out different structural-functional roles. Demonstration of the inner membrane localization of the Bdr proteins and of the differences in nature of the interaction of individual Bdr paralogs with the cell infrastructure is an important step toward defining the functional role of this unique protein family in the genus Borrelia.
Roberts
D M
DM
Department of Microbiology and Immunology, School of Medicine, Medical College of Virginia at Virginia Commonwealth University, Richmond, Virginia 23298-0678, USA.
Theisen
M
M
Marconi
R T
RT
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D015748:Borrelia burgdorferi Group / Q000166:cytology* / Q000235:genetics*; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D007106:Immune Sera; D008193:Lyme Disease / Q000382:microbiology*; D005810:Multigene Family; D012995:Solubility
D001426:Bacterial Proteins; D003902:Detergents; D007106:Immune Sera