부산대학교 도서관

As an enzootic pathogen, the Lyme disease bacterium Borrelia burgdorferi possesses multiple copies of chemotaxis proteins, including two chemotaxis histidine kinases (CHK), CheA1 and CheA2. Our previous study showed that CheA2 is a genuine CHK that is required for chemotaxis; however, the role of CheA1 remains mysterious. This report first compares the structural features that differentiate CheA1 and CheA2 and then provides evidence to show that CheA1 is an atypical CHK that controls the virulence of B. burgdorferi through modulating the stability of RpoS, a key transcriptional regulator of the spirochete. First, microscopic analyses using green-fluorescence-protein (GFP) tags reveal that CheA1 has a unique and dynamic cellular localization. Second, loss-of-function studies indicate that CheA1 is not required for chemotaxis in vitro despite sharing a high sequence and structural similarity to its counterparts from other bacteria. Third, mouse infection studies using needle inoculations show that a deletion mutant of CheA1 (cheA1mut) is able to establish systemic infection in immune-deficient mice but fails to do so in immune-competent mice albeit the mutant can survive at the inoculation site for up to 28 days. Tick and mouse infection studies further demonstrate that CheA1 is dispensable for tick colonization and acquisition but essential for tick transmission. Lastly, mechanistic studies combining immunoblotting, protein turnover, mutagenesis, and RNA-seq analyses reveal that depletion of CheA1 affects RpoS stability, leading to reduced expression of several RpoS-regulated virulence factors (i.e., OspC, BBK32, and DbpA), likely due to dysregulated clpX and lon protease expression. Bulk RNA-seq analysis of infected mouse skin tissues further show that cheA1mut fails to elicit mouse tnf-α, il-10, il-1β, and ccl2 expression, four important cytokines for Lyme disease development and B. burgdorferi transmigration. Collectively, these results reveal a unique role and regulatory mechanism of CheA1 in modulating virulence factor expression and add new insights into understanding the regulatory network of B. burgdorferi. Author summary: Lyme disease is an infectious disease caused by the bacterium Borrelia burgdorferi which is transmitted to humans through the bite of infected blacklegged ticks. It is the most commonly reported tick-borne illness in North America and Europe. B. burgdorferi is a highly invasive bacterium that can swim and penetrate deep into tissues to cause a wide range of disorders including arthritis, meningitis, and migratory musculoskeletal pain. This invasiveness is driven by the ability of the spirochete to sense and respond to its surrounding initiated by signal transduction coupled with a sophisticated array of sensory proteins located at both cell ends. The genome of B. burgdorferi encodes multiple copies of chemotaxis-related genes and the function of many of these genes remains unknown. In this report, we focus on deciphering the role of one of the chemotaxis histidine kinases, CheA1. Our results reveal a novel regulatory contribution of CheA1 to the infectious life cycle and pathogenesis of B. burgdorferi. [ABSTRACT FROM AUTHOR]