Biofilm production is frequently dependent on such environmental factors as cell density and glucose concentration. The Enterococcus faecalis quorum-sensing locus (fsr) increases enterococcal virulence in multiple animal models. To date, fsr has been shown to regulate the transcription of 2 downstream protease genes. We demonstrate that the effect of fsr mutations on biofilm formation, as well as the fsr-mediated catabolite control of biofilm, is mediated via these proteases. The present study provides additional insight into the mechanisms used by E. faecalis to establish nosocomial infection.