Contribution of RpoS to persistence of Borrelia burgdorferi in mammals

It is now universally accepted that the alternative sigma factor RpoS serves as a master regulator for differential gene expression in Bb. Using a novel enrichment RNAseq technique, we established that the RpoS regulon changes dramatically as spirochetes undergo mammalian host adaptation and that the mammalian phase RpoS regulon includes virulence determinants required for early infection. The unexpected discovery that the BosR/RpoS regulon includes vlsE was a clear indicator that RpoS is needed not only to establish infection but also to maintain it. Using a rpoS mutant complemented with an IPTG-inducible rpoS allele, we established that Bb do not survive in mice once RpoS is turned OFF and that persistence involves elements within the RpoS regulatory network functionally unrelated to evasion of adaptive immunity. These findings set the stage for Aims to achieve our overarching objective – elucidating how RpoS-dependent gene regulation perpetuates the mammalian phase of the spirochete's enzootic cycle. In Aim 1, we use our IPTG-inducible system for controlling borrelial gene expression in vivo interrogation individual RpoS-regulated genes in specific milieus and time points throughout the course of murine infection. In Aim 2, we dissect the topological features and promoter elements used by RpoS, BosR and YebC to collaboratively control expression of vlsE and determine the relationship between expression of vlsE and switching in vivo. The proposed experiments will shed new light on how LD spirochetes persist in nature, the ecological substrate for human disease, as well as how they persist in human tissues, giving rise to the chronic, debilitating clinical manifestations that make them a major threat to health and wellbeing. Project Number: 1R21AI187932-01 | Fiscal Year: 2025 | NIH Institute/Center: National Institute of Allergy and Infectious Diseases (NIAID) | Principal Investigator: MELISSA CAIMANO (+1 co-PI) | Institution: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT, FARMINGTON, CT | Award Amount: $457,875 | Activity Code: R21 | Study Section: Bacterial Virulence Study Section [BV] View on NIH RePORTER: https://reporter.nih.gov/project-details/1R21AI18793201