1887

Abstract

The () gene of , the agent of Lyme disease, encodes a surface-exposed lipoprotein. The expression of is tightly regulated and dependent on several environmental factors. In nature, its expression is induced in the tick vector during feeding and maintained during infection of the vertebrate host. The pattern of expression of suggests that it imparts a critical function to the pathogen. A previous study has shown that the expression of is down-regulated in the absence of RpoS, suggesting that the alternative sigma factor may be involved in its expression. A DNA-binding protein has also been shown to specifically recognize a sequence in the 5′ regulatory region of the gene. Therefore, the contribution of these putative determinants to the differential expression of was investigated. The role of RpoS was critically evaluated by genetic complementation of the mutant using a chromosomally targeted copy of the wild-type gene. The results confirm that RpoS is indeed required for the expression of . The role of the upstream DNA-binding site was examined using promoter– transcriptional fusions in a shuttle vector. The DNA-binding site was studied by targeting mutations to an inverted repeat sequence (IRS), the most prominent feature within the binding site, as well as by deletion of the entire sequence upstream of the basal promoter. Quantitative assessment of gene expression demonstrated that neither the IRS nor the sequence upstream of the promoter was essential for expression. Moreover, the expression of the reporter (GFP) appeared to remain RpoS-dependent in all cases, based on the co-expression of GFP and OspC in a subpopulation of spirochaetes and the selective expression of GFP in the stationary phase. Collectively, the data indicate that RpoS is the sole determinant of differential expression in cultured spirochaetes.

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2008-01-01
2024-03-28
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