Role of mprF1 and mprF2 in the pathogenicity of Enterococcus faecalis.
BACKGROUND
Enterococcus faecalis is one of the leading causes of nosocomial infections. Due to its innate and acquired resistance to most antibiotics, identification of new targets for antimicrobial treatment of E. faecalis is a high priority. The multiple peptide resistance factor MprF, which was first described in Staphylococcus aureus, modifies phosphatidylglycerol with lysin and reduces the negative charge of the membrane, thus increasing resistance to cationic antimicrobial peptides. We studied the effect of mprF in E. faecalis regarding influence on bacterial physiology and virulence.


RESULTS
Two putative mprF paralogs (mprF1 and mprF2) were identified in E. faecalis by BLAST search using the well-described S. aureus gene as a lead. Two deletion mutants in E. faecalis 12030 were created by homologous recombination. Analysis of both mutants by thin-layer chromatography showed that inactivation of mprF2 abolishes the synthesis of three distinct amino-phosphatidylglycerols (PGs). In contrast, deletion of mprF1 did not interfere with the biosynthesis of amino-PG. Inactivation of mprF2 increased susceptibility against several antimicrobial peptides and resulted in a 42% increased biofilm formation compared to wild-type mprF. However, resistance to opsonic killing was increased in the mutant, while virulence in a mouse bacteremia model was unchanged.


CONCLUSIONS
Our data suggest that only mprF2 is involved in the aminoacylation of PG in enterococci, and is probably responsible for synthesis of Lys-PG, Ala-PG, and Arg-PG, while mprF1 does not seem to have a role in aminoacylation. As in other Gram-positive pathogens, aminoacylation through MprF2 increases resistance against cationic antimicrobial peptides. Unlike mprF found in other bacteria, mprF2 does not seem to be a major virulence factor in enterococci.
Division of Infectious Diseases, Department of Medicine, University Hospital Freiburg, Freiburg, Germany.
Yinyin
Y
Bao
Tuerkan
T
Sakinc
Diana
D
Laverde
Dominique
D
Wobser
Abdellah
A
Benachour
Christian
C
Theilacker
Axel
A
Hartke
Johannes
J
Huebner
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D018441:Biofilms / Q000254:growth & development; D024881:Drug Resistance, Bacterial / Q000235:genetics; D013293:Enterococcus faecalis / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D017353:Gene Deletion; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology*; D051379:Mice; D010743:Phospholipids / Q000378:metabolism; D017422:Sequence Analysis, DNA; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism
D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins; D010743:Phospholipids; D037521:Virulence Factors