1887

Abstract

The resuscitation-promoting factors of are hydrolytic enzymes, which are required for resuscitation of dormant cells. RpfB, a peptidoglycan remodelling enzyme similar to the lytic transglycosylase of , is required for reactivation of from chronic infection , underscoring the need to understand its transcriptional regulation. Here, we identified the transcriptional and translational start points of , and suggested from promoter-driven GFP expression and transcription assays that its transcription possibly occurs in a SigB-dependent manner. We further demonstrated that transcription is regulated by MtrA – the response regulator of the essential two-component system MtrAB. Association of MtrA with the promoter region was confirmed by chromatin immunoprecipitation analysis. Electrophoretic mobility shift assays (EMSAs) revealed a loose direct repeat sequence associated with MtrA binding. Binding of MtrA was enhanced upon phosphorylation. MtrA could be pulled down from lysates of using a biotinylated DNA fragment encompassing the MtrA-binding site on the promoter, confirming that MtrA binds to the promoter. Enhanced GFP fluorescence driven by the promoter, upon deletion of the MtrA-binding site, and repression of expression, upon overexpression of MtrA, suggested that MtrA functions as a repressor of transcription. This was corroborated by EMSAs showing diminished association of RNA polymerase (RNAP) with the promoter in the presence of MtrA. transcription assays confirmed that MtrA inhibits RNAP-driven transcription.

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/content/journal/micro/10.1099/mic.0.000087
2015-06-01
2022-01-26
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