Gram-negative quorum sensing signalling enhances biofilm formation and virulence traits in gram-positive pathogen Enterococcus faecalis

J Oral Microbiol. 2023 May 11;15(1):2208901. doi: 10.1080/20002297.2023.2208901. eCollection 2023.

Abstract

Acyl-homoserine lactones (AHLs) are typical quorum-sensing molecules of gram-negative bacteria. Recent evidence suggests that AHLs may also affect gram-positives, although knowledge of these interactions remains scarce. Here, we assessed the effect of AHLs on biofilm formation and transcriptional regulations in the gram-positive Enterococcus faecalis. Five E. faecalis strains were investigated herein. Crystal violet was employed to quantify the biomass formed, and confocal microscopy in combination with SYTO9/PI allowed the visualisation of biofilms' structure. The differential expression of 10 genes involved in quorum-sensing, biofilm formation and stress responses was evaluated using reverse-transcription-qPCR. The AHL exposure significantly increased biofilm production in strain ATCC 29212 and two isolates from infected dental roots, UmID4 and UmID5. In strains ATCC 29212 and UmID7, AHLs up-regulated the quorum-sensing genes (fsrC, cylA), the adhesins ace, efaA and asa1, together with the glycosyltransferase epaQ. In strain UmID7, AHL exposure additionally up-regulated two membrane-stress response genes (σV, groEL) associated with increased stress-tolerance and virulence. Altogether, our results demonstrate that AHLs promote biofilm formation and up-regulate a transcriptional network involved in virulence and stress tolerance in several E. faecalis strains. These data provide yet-unreported insights into E. faecalis biofilm responses to AHLs, a family of molecules long-considered the monopole of gram-negative signalling.

Keywords: Enterococcus faecalis; Fsrc; acyl-homoserine lactones; biofilm formation; cell-to-cell communication; endodontic infection; gene expression; quorum sensing; reverse-transcription quantitative PCR; virulence.

Grants and funding

This work was supported by a doctoral fellowship from the Regional Government of Galicia, Spain [ED481A-2019/194] (AP); University of Geneva Institutional Funds for Senior Clinical Researchers (DM); a PDTS Project from the Instituto de Salud Carlos III, Spain [DTS21/00015] (AO); Karolinska Institutet Strategic Funds (GNB); KI/SLL Styrgruppen för Odontologisk Forskning (SOF) [Dnr. 4- 823/2019] (GNB); KI/SLL Styrgruppen för Odontologisk Forskning (SOF) [FoUI-978687] (GNB); Vetenskapsrådet [Project n°2022-1014] (GNB).