Regulatory roles of the AraC family transcription factor yeaM in the virulence and biofilm formation of Salmonella Typhimurium

Abstract

Salmonella Typhimurium (S. typhimurium) is a significant zoonotic pathogen responsible for gastroenteritis and severe systemic infections in various hosts. The AraC family transcription factors are key gene expression regulators in prokaryotes, essential for bacterial adaptation to the environment and virulence. Despite their importance, the role of yeaM, a member of this family in S. typhimurium, remains unexplored.

To elucidate yeaM regulatory function in virulence and biofilm formation, we engineered mutant and complementary strains of the yeaM gene using homologous recombination. We assessed their capabilities in biofilm formation under different conditions, macrophage adherence and invasion, and virulence in mice. Additionally, we identified potential target genes regulated by yeaM through transcriptome sequencing and confirmed these findings using an electrophoretic mobility shift assay (EMSA) and a dual-luciferase reporter assay.

Our results demonstrate that, compared to the parental strain SL1344 and the complemented strain CΔyeaM, the ΔyeaM strain exhibited significantly enhanced biofilm formation, increased invasion of mouse intestinal epithelial cells, enhanced intracellular proliferation within macrophages, and elevated induction of macrophage apoptosis. Furthermore, the ΔyeaM deletion strain displayed significantly increased virulence in mice and enhanced proliferation in milk. Transcriptome analysis revealed that S. typhimurium pathogenicity island 4 (SPI4) genes (siiA, siiB, siiC, siiD, siiF, and siiE) were significantly upregulated following the deletion of the yeaM gene. EMSA and dual-luciferase reporter assays further showed that the yeaM protein can bind to the promoter of the siiA gene and suppress its expression, thereby modulating the biofilm formation and virulence of S. typhimurium.