Fsr-independent production of protease(s) may explain the lack of attenuation of an Enterococcus faecalis fsr mutant versus a gelE-sprE mutant in induction of endocarditis.
An Enterococcus faecalis gelE insertion disruption mutant (TX5128), which produces neither gelatinase (GelE) nor the cotranscribed (in the wild type) serine protease (SprE), was found to be attenuated in a rat endocarditis model with a significant decrease in the endocarditis induction rate versus wild-type E. faecalis OG1RF (GelE(+), SprE(+)). TX5266, which has a nonpolar deletion in fsrB and, like TX5128, is phenotypically GelE(-) under usual conditions, was also studied; fsrB is a homologue of agrB of staphylococci and participates in regulation of gelE-sprE expression. Unexpectedly, TX5266 approximated wild-type OG1RF in the endocarditis model and was significantly less attenuated than TX5128. This is in contrast to other models which have found fsr mutants to be as or more attenuated than TX5128. Further study found that the fsrB mutant produced very low levels of gelatinase activity after prolonged incubation in vitro versus no gelatinase activity with TX5128 and did not show the extensive chaining characteristic of TX5128. Reverse transcription-PCR confirmed that gelE was expressed in TX5266 at a very low level versus wild-type OG1RF and was not expressed at all in TX5128. Possible explanations for the increased induction of endocarditis by TX5266 versus TX5128 include the production of low levels of protease(s) or some other effect(s) of the inactivation of the E. faecalis fsr regulator. The equivalent ability of OG1RF and its fsr mutant to initiate endocarditis may explain why we did not find naturally occurring fsr mutants, which account for ca. 35% of E. faecalis isolates, unrepresented in endocarditis versus fecal isolates.
Singh
Kavindra V
KV
Division of Infectious Diseases, Center for the Study of Emerging and Re-emerging Pathogens, University of Texas Medical School at Houston, 6431 Fannin, 2.112 MSB, Houston, TX 77030, USA.
Nallapareddy
Sreedhar R
SR
Nannini
Esteban C
EC
Murray
Barbara E
BE
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004697:Endocarditis, Bacterial / Q000201:enzymology* / Q000382:microbiology; D013293:Enterococcus faecalis / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D018093:Gelatinases / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D009154:Mutation; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism
D001426:Bacterial Proteins; C409032:FsrB protein, Enterococcus; D012697:Serine Endopeptidases; D018093:Gelatinases