Residue L193P Mutant of RpoS Affects Its Activity During Biofilm Formation in Salmonella Pullorum.

The role of alternative sigma factor RpoS in regulating biofilm formation may differ in various Salmonella Pullorum strains. In this study, the biofilm-forming ability of two Salmonella Pullorum strains S6702 and S11923-3 were compared. The biofilm forming ability of S11923-3 was much stronger than that of S6702. After knocking out the rpoS gene, S11923-3ΔrpoS had significantly reduced biofilm while S6702ΔrpoS demonstrated similar biofilm compared with each parent strain. The analysis of RpoS sequences indicated two amino acid substitutions (L193P and R293C) between S6702 and S11923-3 RpoS. A complementation study confirmed that the expression of S11923-3 RpoS rather than S6702 RpoS could restore the biofilm-forming ability of ΔrpoS strains and the L193P mutation contributed to the restoration of the biofilm-forming ability. Further study indicated that RpoS with the L193P mutant had significantly improved expression level and binding activity to RNAP and csgD gene promoter, which increased the efficacy of the csgD gene promoter and biofilm-forming ability. Therefore, the L193P mutation of RpoS is critical for stronger biofilm formation of Salmonella Pullorum.

Contribution of the csgA and bcsA genes to Salmonella enterica serovar Pullorum biofilm formation and virulence.

Salmonella biofilm formation is important to environmental stress resistance and virulence. However, the roles of the csgA and bcsA genes, which affect curli protein and cellulose production, respectively, in Salmonella enterica serovar Pullorum, are unknown. Here we constructed deletions in the csgA and bcsA genes in S. enterica serovar Pullorum strain S6702 and evaluated several aspects of biofilm formation and virulence. ΔcsgA showed decreased production of curli fimbriae, while ΔbcsA had reduced cellulose production. Both mutants had a reduced ability to form biofilms. ΔcsgA was reduced in adhesion and invasion to HeLa cells and exhibited decreased intracellular proliferation in HD11 macrophages. ΔbcsA exhibited increased proliferation in HD11 cells and replicated better in chicken spleens, as compared to the wild-type strain. ΔcsgA virulence was attenuated in assays involving oral challenge of one-day-old chickens.