Listeria monocytogenes σH Contributes to Expression of Competence Genes and Intracellular Growth.

Abstract

The alternative sigma factor σ(H)has two functions in Gram-positive bacteria: it regulates sporulation and the development of genetic competence. Listeria monocytogenes is a nonsporulating species in which competence has not yet been detected. Nevertheless, the main competence regulators and a series of orthologous genes that form the competence machinery are present in its genome; some of the competence genes play a role in optimal phagosomal escape. In this study, strains overexpressing σ(H) and strains with a σ(H) deletion were used to elucidate the contribution of σ(H) to the expression of the competence machinery genes inL. monocytogenes Gene expression analysis showed that σ(H) is, indeed, involved in comG and come regulation. Unexpectedly, we observed a unique regulation scheme in which σ(H) and the transcription factor ComK were involved. Population-level analysis showed that even with the overexpression of both factors, only a fraction of the cells expressed the competence machinery genes. Although we could not detect competence, σ(H) was crucial for phagosomal escape, which implies that this alternative sigma factor has specifically evolved to regulate the L. monocytogenes intracellular life cycle. Listeria monocytogenes can be an intracellular pathogen capable of causing serious infections in humans and animal species. Recently, the competence machinery genes were described as being necessary for optimal phagosomal escape, in which the transcription factor ComK plays an important role. On the other hand, our previous phylogenetic analysis suggested that the alternative sigma factor σ(H) might play a role in the regulation of competence genes. The present study shows that some of the competence genes belong to the σ(H) regulon and, importantly, that σ(H) is essential for intracellular growth, implying a unique physiological role of σ(H) among Firmicutes.