Article Figures & Data
Figures
- Fig. 1.
Open reading frames and their transcripts ingelE flanking regions. (A) Open reading frames. The solid line represents the chromosome, and the genes and open reading frames are indicated by boxes in different shades. The orientation of the genes and open reading frames are indicated by arrows. (B) Summary of the Northern blot results using RNA from different strains and different gene probes. −, No signal in Northern blot analysis.
- Fig. 2.
Sequence alignment of FsrA, FsrB, and FsrC with AgrA, AgrB, and AgrC from S. aureus. Identical amino acids are indicated by a symbol as “∣”, and similar amino acids are indicated by a symbol as “:” or “.” between them. Conserved residues in H, N, and G2 blocks of histidine protein kinases and conserved Asp and Lys residues in response regulators of bacterial two-component systems (20) are boxed. The sequences fromS. aureus were as follows: AgrA, accession number M21854 (14); AgrB, accession number AF001782 (5); and AgrC, accession number AF001783 (5).
- Fig. 3.
Gelatinase and serine protease activities infsr and sprE mutants. These pictures are the negative images of gelatin and casein zymogram gels. (A) Gelatinase activity of mutants on gelatin zymogram gel. (B) Serine protease activity of mutants on casein zymogram gel. Lanes: o1, mutant with disruption of orf1; o, OG1RF; a, mutant with disruption offsrA; b, mutant with disruption of fsrB; c, mutant with disruption of fsrC; e, mutant with disruption ofgelE; s, mutant with disruption of sprE.
- Fig. 4.
Northern blot analysis of fsr andsprE mutants. (A, B, C, and D) Northern blots of mutants using fsrB, fsrC, gelE, andsprE probes, respectively. Lanes: o1, mutant with disruption of orf1; o, OG1RF; a, mutant with disruption offsrA; b, mutant with disruption of fsrB; c, mutant with disruption of fsrC; s, mutant with disruption ofsprE.
- Fig. 5.
Kaplan-Meier survival plots of wild-type OG1RF andfsr and sprE mutants in the mouse peritonitis model. (A) Kaplan-Meier survival plots of OG1RF, the fsrAmutant (TX5240) and an fsrC revertant. (B) OG1RF and thefsrB mutant (TX5241). (C) OG1RF and the sprEmutant (TX5243). Twelve mice were tested with each inoculum of each of the strains shown. The P value refers to mutant versus OG1RF at the same or a smaller inoculum (▴ versus ▵ or ■ versus □).
Tables
- Table 1.
Bacterial strains and plasmids
Strain or plasmid Relevant genotype or phenotype Source or reference Strain OG1RF Gelatinase and serine protease production positive/(Gel+, Spr+) 10 TX5128 gelE transposon mini-γδ insertion mutant of OG1RF, Gel−, Spr−, Kanr 19 TX5240 fsrA insertion mutant of OG1RF, Gel−, Spr−, Kanr This study TX5241 fsrB insertion mutant of OG1RF, Gel−, Spr−, Kanr This study TX5242 fsrC insertion mutant of OG1RF, Gel−, Spr−, Kanr This study TX5243 sprE insertion mutant of OG1RF, Gel+, Spr−, Kanr This study TX5248 orf1 insertion mutant of OG1RF, Gel+, Spr+, Kanr This study TX5244 TX5240 harboring plasmid TEX5249, Gel+, Spr+, Kanr Emr This study TX5245 TX5241 harboring plasmid TEX5249, Gel+, Spr+, Kanr Emr This study TX5246 TX5242 harboring plasmid TEX5249, Gel+, Spr+, Kanr Emr This study TX5247 TX5128 harboring plasmid TEX5249, Gel−, Spr−, Kanr Emr This study DH5α E. coli host strain for routine cloning Stratagene TX5249 DH5α(pTEX5249) This study Plasmids pBluescript SK(−) Cloning vector, Ampr Stratagene pTEX4577 Suicide vector for single cross-over mutagenesis in E. faecalis, Kanr 19 pAT18 Shuttle vector for E. coli and E. faecalis, Emr 23 pTEX5249 pTEX5249 was constructed by cloning a 6-kb PstI/BglII fragment containing fsrA, fsrB, and fsrC into the shuttle vector pAT18, Emr This study
