Actinobacillus pleuropneumoniae two-component system QseB/QseC regulates the transcription of PilM, an important determinant of bacterial adherence and virulence

- Generation of A. pleuropneumoniae QseB/QseC deletion mutant ΔqseBC and PilM mutant ΔpilM.
- 44 differently expressed genes in ΔqseBC were identified by microarray.
- Direct regulation of PilM by QseB was confirmed by EMSA.
- PilM mutation leads to reduced adherence ability to SJPL cells.
- PilM contributes to the pathogenesis in pigs and is an important virulence determinant.

QseB/QseC is one of the five predicted two-component systems (TCSs) in Actinobacillus pleuropneumoniae. To understand the roles of this TCS in A. pleuropneumoniae, a markerless gene-deletion mutant ΔqseBC was constructed. Differentially expressed (DE) genes in ΔqseBC were filtered by microarray analysis. A total of 44 DE genes were found to be regulated by QseB/QseC system. The transcriptional profile of A. pleuropneumoniae ΔqseBC was compared with that of ΔluxS and catecholamine (CA) stimulations, 13 genes regulated by QseB/QseC were found also regulated by LuxS, and 3 Qse-regulons were co-regulated by CA stimulations, respectively. Binding of QseB to the promoters of three regulons (pilM, glpK and hugZ), which were co-regulated by QseB/QseC and LuxS, was evaluated by electrophoretic mobility-shift assay. Results indicated that pilM was directly regulated by phosphorylated-QseB. Then the pilM deletion mutant ΔpilM was constructed and characterized. Data presented here revealed that adherence ability of ΔpilM to St. Jude porcine lung cells was significantly decreased, and ΔpilM exhibited reduced virulence in pigs, suggesting PilM contributes to the process of A. pleuropneumoniae infection.