Characterization of a negative regulator of exopolysaccharide production by the plant-pathogenic bacterium Pseudomonas solanacearum.
Wild-type strains of the bacterial wilt pathogen Pseudomonas solanacearum exhibit reduced exopolysaccharide production and virulence when transformed with plasmids carrying the epsR locus. To understand the function of epsR, we used mutagenesis and DNA sequencing to identify the gene responsible for the shutoff of exopolysaccharide production. The epsR gene encodes a 236-amino-acid polypeptide that, based on polypeptide sequence homology, has significant similarity to other proteins of the luxR family of environmentally responsive, two-component regulatory systems. When a mutated copy of the epsR gene was marker-exchanged into the wild-type P. solanacearum chromosome, however, we observed no effect on growth in culture or on exopolysaccharide production. This suggests that the EpsR phenotype becomes apparent only via overproduction of the EpsR protein. By means of an antiserum directed against the EpsR protein, we detected the overproduction of EpsR in cell lysates of a strain of P. solanacearum harboring a multicopy plasmid with an active epsR gene but not in one harboring the same plasmid with a mutated epsR gene.
Kao
C C
CC
Department of Plant Pathology, University of Wisconsin, Madison 53706.
Gosti
F
F
Huang
Y
Y
Sequeira
L
L
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004926:Escherichia coli; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D010944:Plants / Q000382:microbiology*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis*; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D012097:Repressor Proteins; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators; D014157:Transcription Factors; D014774:Virulence
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D011135:Polysaccharides, Bacterial; D012097:Repressor Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C087016:epsR protein, Ralstonia solanacearum; C064778:LuxR autoinducer binding proteins