Microarray analysis of quorum-sensing-regulated genes in Porphyromonas gingivalis.
Quorum sensing is a phenomenon defined as gene regulation in response to cell density that regulates various functions in bacteria. The periodontopathogen Porphyromonas gingivalis possesses a luxS gene homologue that may encode a quorum-sensing system. In order to identify genes of P. gingivalis that are regulated by luxS, gene expression analysis was done using microarrays and RNA samples from the W83 wild-type strain and an isogenic luxS mutant, LY2001. The results indicated that 17 open reading frames (ORFs) in LY2001 are upregulated and two are downregulated. Real-time PCR was done to confirm the microarray results. Among the upregulated ORFs is a group of stress-related genes, including htrA, clpB, groEL, dnaK, and the F subunit of alkyl hydroperoxide reductase. This suggested that luxS is involved in stress gene regulation in P. gingivalis. Stress response experiments, including high-temperature survival, resistance to hydrogen peroxide (H2O2), and survival during exposure to low and high pH, were performed on the P. gingivalis wild-type and LY2001 strains. LY2001 had a significantly higher survival rate than did W83 when stressed at 50 degrees C. No difference was found at pH 5, but LY2001 had increased survival compared to W83 at pH 9. LY2001 also survived better than W83 when stressed with 0.35 mM H2O2. These results suggest that luxS might be involved in promoting survival of P. gingivalis in the host by regulating its response to host-induced stresses such as temperature, H2O2, and pH.
Yuan
Lihui
L
Department of Oral Biology and Center for Molecular Microbiology, College of Dentistry, University of Florida, P.O. Box 100424, Gainesville, Florida 32610-0424, USA.
Hillman
Jeffrey D
JD
Progulske-Fox
Ann
A
D001426:Bacterial Proteins / Q000502:physiology*; D013437:Carbon-Sulfur Lyases; D015964:Gene Expression Regulation, Bacterial; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008163:Luminescent Measurements; D020411:Oligonucleotide Array Sequence Analysis; D016966:Porphyromonas gingivalis / Q000235:genetics* / Q000254:growth & development; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013696:Temperature
D001426:Bacterial Proteins; D006861:Hydrogen Peroxide; D013437:Carbon-Sulfur Lyases; C117753:LuxS protein, Bacteria