Proteomic analysis of quorum sensing-dependent proteins in Burkholderia glumae.
Burkholderia glumae, the causal agent of bacterial rice grain rot, utilizes quorum sensing (QS) systems that rely on N-octanoyl homoserine lactone (synthesized by TofI) and its cognate receptor TofR to activate toxoflavin biosynthesis genes and an IclR-type transcriptional regulator gene, qsmR. Since QS is essential for B. glumae pathogenicity, we analyzed the QS-dependent proteome by 2-dimensional gel electrophoresis. A total of 79 proteins, including previously known QS-dependent proteins, were differentially expressed between the wild-type BGR1 and the tofI mutant BGS2 strains. Among this set, 59 proteins were found in the extracellular fraction, and 20 were cytoplasmic. Thirty-four proteins, including lipase and proteases, were secreted through the type II secretion system (T2SS). Real-time RT-PCR analysis showed that the corresponding genes of the 49 extracellular and 13 intracellular proteins are regulated by QS at the transcriptional level. The T2SS, encoded by 12 general secretion pathway (gsp) genes with 3 independent transcriptional units, was controlled by QS. beta-Glucuronidase activity analysis of gsp::Tn3-gusA gene fusions and electrophoretic mobility shift assays revealed that the expression of gsp genes is directly regulated by QsmR. T2SS-defective mutants exhibited reduced virulence, indicating that the T2SS-dependent extracellular proteins play important roles in B. glumae virulence.
Goo
Eunhye
E
Department of Agricultural Biotechnology, Seoul National University, Seoul, Republic of Korea.
Kang
Yongsung
Y
Kim
Hongsup
H
Hwang
Ingyu
I
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism; D019117:Burkholderia / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics / Q000502:physiology; D012275:Oryza / Q000382:microbiology; D040901:Proteomics / Q000379:methods*; D053038:Quorum Sensing / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055571:Secretory Pathway; D053719:Tandem Mass Spectrometry; D037521:Virulence Factors
D001426:Bacterial Proteins; D037521:Virulence Factors