Dual signaling functions of the hybrid sensor kinase RpfC of Xanthomonas campestris involve either phosphorelay or receiver domain-protein interaction.
The hybrid sensor kinase RpfC positively regulates the expression of a range of virulent genes and negatively modulates the synthesis of the quorum sensing signal diffusible signal factor (DSF) in Xanthomonas campestris. Three conserved amino acid residues of RpfC implicated in phosphorelay (His(198) in the histidine kinase domain, Asp(512) in the receiver domain, and His(657) in the histidine phosphotransfer domain) were essential for activation of the production of extracellular enzymes and extracellular polysaccharide (EPS) virulence factors but were not essential for repression of DSF biosynthesis. Domain deletion and subsequent in trans expression analysis revealed that the receiver domain of RpfC alone was sufficient to repress DSF overproduction in an rpfC deletion mutant. Further deletion and alanine scanning mutagenesis analyses identified a peptide of 107 amino acids and three amino acid residues (Gln(496), Glu(504), and Ile(552)) involved in modulating DSF production. Co-immunoprecipitation and far Western blot analyses suggested an interaction between the receiver domain and RpfF, the enzyme involved in DSF biosynthesis. These data support a model in which RpfC modulates two different functions (virulence factor synthesis and DSF synthesis) by utilization of a conserved phosphorelay system and a novel domain-specific protein-protein interaction mechanism, respectively. This latter mechanism represents an added dimension to conventional two-component signaling paradigms.
He
Ya-Wen
YW
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Singapore 138673.
Wang
Chao
C
Zhou
Lian
L
Song
Haiwei
H
Dow
J Maxwell
JM
Zhang
Lian-Hui
LH
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis; D011485:Protein Binding; D015398:Signal Transduction; D016959:Xanthomonas campestris / Q000201:enzymology* / Q000235:genetics
D000596:Amino Acids; D001426:Bacterial Proteins; D011135:Polysaccharides, Bacterial; C102500:rpfC protein, Xanthomonas