Gac is a transcriptional repressor of the Lyme disease spirochete’s OspC virulence-associated surface protein

Abstract

AbstractThe OspC outer-surface lipoprotein is essential for the Lyme disease spirochete’s initial phase of vertebrate infection. Bacteria within the midguts of unfed ticks do not express OspC, but produce high levels when ticks begin to ingest blood. Lyme disease spirochetes cease production of OspC within 1-2 weeks of vertebrate infection, and bacteria that fail to downregulate OspC are cleared by host antibodies. Thus, tight regulation of OspC levels is critical for survival of Lyme borreliae, and therefore an attractive target for development of novel treatment strategies. Previous studies determined that a DNA region 5’ of theospCpromoter, theospCoperator, is required for control of OspC production. Hypothesizing that theospCoperator may bind a regulatory factor, DNA affinity pulldown was performed, and identified binding by the Gac protein. Gac is encoded by the C-terminal domain of thegyrAopen reading frame, from an internal promoter, ribosome-binding site, and initiation codon. Our analyses determined that Gac exhibits a greater affinity forospCoperator and promoter DNAs than for other tested borrelial sequences. In vitro and in vivo analyses demonstrated that Gac is a transcriptional repressor ofospC. These results constitute a substantial advance to our understanding the mechanisms by which the Lyme disease spirochete controls production of OspC.ImportanceBorrelia burgdorferi(sensu lato) requires its surface-exposed OspC protein in order to establish infection of humans and other vertebrate hosts. Bacteria that either do not produce OspC during transmission, or fail to repress OspC after infection is established, are rapidly cleared by the host. Herein, we identified a borrelial protein, Gac, that exhibits preferential affinity to theospCpromoter and 5’ adjacent DNA. A combination of biochemical analyses and investigations of genetically-manipulated bacteria demonstrated that Gac is a transcriptional repressor ofospC. This is a substantial advance toward understanding how the Lyme disease spirochete controls production of the essential OspC virulence factor, and identifies a novel target for preventative and curative therapies.