Refinement of the Listeria monocytogenes σB regulon through quantitative proteomic analysis

Abstract

σ(B) is an alternative σ factor that regulates stress response and virulence genes in the foodborne pathogen Listeria monocytogenes. To gain further insight into σ(B)-dependent regulatory mechanisms in L. monocytogenes, we (i) performed quantitative proteomic comparisons between the L. monocytogenes parent strain 10403S and an isogenic ΔsigB mutant and (ii) conducted a meta-analysis of published microarray studies on the 10403S σ(B) regulon. A total of 134 genes were found to be significantly positively regulated by σ(B) at the transcriptomic level with >75 % of these genes preceded by putative σ(B)-dependent promoters; 21 of these 134 genes were also found to be positively regulated by σ(B) through proteomics. In addition, 15 proteins were only found to be positively regulated by σ(B) through proteomics analyses, including Lmo1349, a putative glycine cleavage system protein. The lmo1349 gene is preceded by a 5' UTR that functions as a glycine riboswitch, which suggests regulation of glycine metabolism by σ(B) in L. monocytogenes. Herein, we propose a model where σ(B) upregulates pathways that facilitate biosynthesis and uptake of glycine, which may then activate this riboswitch. Our data also (i) identified a number of σ(B)-dependent proteins that appear to be encoded by genes that are co-regulated by multiple transcriptional regulators, in particular PrfA, and (ii) found σ(B)-dependent genes and proteins to be overrepresented in the 'energy metabolism' role category, highlighting contributions of the σ(B) regulon to L. monocytogenes energy metabolism as well as a role of PrfA and σ(B) interaction in regulating aspects of energy metabolism in L. monocytogenes.