Homologous response regulators KvgA, KvbA and KvhR regulate the synthesis of capsular polysaccharide in Klebsiella pneumoniae CG43 in a coordinated manner

Abstract

On the basis of phenotypic analysis, the Klebsiella pneumoniae CG43 derived mutants with deletions of the gene encoding respectively the response regulators KvgA, KvhA, and KvhR were classified into two groups. Group I bacteria carrying either kvgA(-) or kvhR(-) exhibited less mucoidy, lower level of capsular polysaccharide (CPS) synthesis and higher LD50 than the parental strain. No apparent change of the group II, including kvhA(-) and hvhA(-)kvhR(-) mutants, was observed. However, the mucoidy of kvhA(-)kvhR(-) mutant was found to be diminished after introducing into a kvhA(-) expressing plasmid. Via promoter-lacZ fusion analysis, kvhA deletion was found to reduce kvhR expression. A regulatory role of KvhA for the expression of kvhR was supported further by EMSA showing a specific binding of KvhA to the putative promoter of kvhR. The promoter activity measurement and EMSA also revealed that KvgA acted as an autoregulator and an activator for the expression of kvhAS and kvhR. In addition, deletion of kvgA suppressed slightly the promoter activity of the cps-orf16-17, and the expression of all three cps transcripts orf1-2, orf3-15, and orf16-17 were reduced in the kvhR(-) mutant. These suggest that the three homologous response regulators interact to control, in coordination, the bacterial cps expression.