Transcriptional control of the H-NS antagonists LeuO and RcsB-BglJ in Escherichia coli

The bacterial nucleoid-associated protein (NAP) H-NS is involved in the organization and compaction of the bacterial chromatin and acts as a global respressor, mainly of genes that have been acquired by horizontal gene transfer and that are related to stress responses and pathogenicity. Binding of H-NS to the DNA and formation of a nucleoprotein complex at promoter regions leads to repression. This repressor effect of H-NS can be antagonized by gene-specific transcription factors (H-NS antagonists) that activate transcription of H NS-repressed genes by competing with H-NS for binding or by disturbing formation of the nucleoprotein complex. Two examples of such H NS antagonists are the LysR-type transcription factor LeuO and the FixJ/NarL-type transcription factor heterodimer RcsB-BglJ. LeuO is a pleiotropic regulator of stress responses and virulence determinants. RcsB-BglJ activates transcription of the H NS-repressed bgl (aryl-β,D-glucoside) operon. In this work, novel targets of RcsB-BglJ were identified in Escherichia coli by microarray analyses. The results suggest that heterodimerization of RcsB and BglJ is essential for regulation. Further, in addition to genes related to unknown or predicted function in the membrane the leuO gene was identified as a target gene. Detailed analysis of transcriptional regulation of leuO demonstrated that RcsB-BglJ strongly activates transcription of leuO by binding proximal to a newly mapped leuO promoter. Thus RcsB-BglJ antagonizes repression of leuO by H-NS and the H-NS-like protein StpA. Additional data presented here show that LeuO negatively autoregulates its own expression and inhibits activation of leuO by RcsB-BglJ. Regulation of leuO by RcsB-BglJ and autoregulation by LeuO, as shown here, as well as activation of bglJ by LeuO, as published previously, indicates a feedback control mechanism of two global transcriptional regulators and H-NS antagonists.This feedback regulation may ensure turn on of their expression in response to specific environmental signals. Screens to search for novel regulators or upstream signals were performed by transposon mutagenesis and by using a genomic expression library. These screens indicate that additional factors may be involved in the regulation of this leuO-bglJ feedback loop