During the previous funding period, we made significant advances in the discovery and characterization of the lipid rafts of Borrelia burgdorferi, the spirochetal agent of Lyme disease. Cholesterol glycolipids form lipid rafts in both in vitro and host-derived spirochetes and that these domains have all the hallmarks of eukaryotic lipid rafts. We further demonstrated that the lipid rafts of B. burgdorferi shift their morphology at different temperatures and have a selective presence of lipoproteins, proteases and their substrates, sensing molecules, and homologs of eukaryotic lipid raft markers. Lipid rafts are dynamic structures with shifting protein cargo that define their biological functions. Fo this renewal application, we will expand our investigations into the dynamic features of the lipid rafts and into their function. The overall hypothesis is that the protein content as well the proten concentrations of lipid rafts change in response to the different conditions that the Borrelia moves through in its vector (tick) - host (mammal) stages, and that these changes influence the adaptation mechanisms required by the organisms to complete their life-cycle. To test this hypothesis, this renewal application will address the following Specific Aims: 1. To characterize the proteomes of the lipid rafts of Borrelia under different biological conditions, and 2. To probe the mechanisms of dynamic change and life-cycle adaptation by studying the contributions of unique elements of the proteome. A number of novel biophysical, imaging and biochemical approaches will provide the results to demonstrate that the Borrelia lipid rafts have important biological functions. Cholesterol as a component of bacterial membranes is being reported in an enlarging number of bacterial pathogens thereby increasing the significance of this proposal, as it is clear that cholesterol utilization is not an isolated feature of the Borreliae.
Lyme disease is increasing in incidence in many parts of the world, including the United States. This chronic spirochetosis is caused by Borrelia burgdorferi. This spirochete has some characteristics that make it unique. Among them, is its ability to incorporate cholesterol onto its membranes, which is something that only some bacteria can accomplish. In this application, we will study the proteins that are associated with the cholestero lipids (collectively known as lipid rafts). This will help in the understanding how these structure assist the spirochete in sensing whether it is in a tick or in a mammal, and how it can adapt to these two fundamentally different environments.
|Coleman, James L; Toledo, Alvaro; Benach, Jorge L (2016) Borrelia burgdorferiâ€…HtrA: evidence for twofold proteolysis of outer membrane protein p66. Mol Microbiol 99:135-50|
|MonzÃ³n, Javier D; Atkinson, Elizabeth G; Henn, Brenna M et al. (2016) Population and Evolutionary Genomics of Amblyomma americanum, an Expanding Arthropod Disease Vector. Genome Biol Evol 8:1351-60|
|Katona, Laura I (2015) The Fur homologue BosR requires Arg39 to activate rpoS transcription in Borrelia burgdorferi and thereby direct spirochaete infection in mice. Microbiology 161:2243-55|
|Toledo, Alvaro; PÃ©rez, Alberto; Coleman, James L et al. (2015) The lipid raft proteome of Borrelia burgdorferi. Proteomics 15:3662-75|
|Toledo, Alvaro; MonzÃ³n, Javier D; Coleman, James L et al. (2015) Hypercholesterolemia and ApoE deficiency result in severe infection with Lyme disease and relapsing-fever Borrelia. Proc Natl Acad Sci U S A 112:5491-6|
|Toledo, Alvaro; Crowley, Jameson T; Coleman, James L et al. (2014) Selective association of outer surface lipoproteins with the lipid rafts of Borrelia burgdorferi. MBio 5:e00899-14|
|Coleman, James L; Crowley, Jameson T; Toledo, Alvaro M et al. (2013) The HtrA protease of Borrelia burgdorferi degrades outer membrane protein BmpD and chemotaxis phosphatase CheX. Mol Microbiol 88:619-33|
|LaRocca, Timothy J; Pathak, Priyadarshini; Chiantia, Salvatore et al. (2013) Proving lipid rafts exist: membrane domains in the prokaryote Borrelia burgdorferi have the same properties as eukaryotic lipid rafts. PLoS Pathog 9:e1003353|
|Crowley, Jameson T; Toledo, Alvaro M; LaRocca, Timothy J et al. (2013) Lipid exchange between Borrelia burgdorferi and host cells. PLoS Pathog 9:e1003109|
|Garcia-Monco, Juan C; Benach, Jorge L (2013) A disconnect between the neurospirochetoses in humans and rodent models of disease. PLoS Pathog 9:e1003288|
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