Pheromone cCF10 Enhances Persister Formation in Enterococcus faecalis via Transcriptomic Changes

Bacterial persistence, a non-heritable high-antibiotic-tolerance phenotype, is a key driver of recurrent clinical infections and antibiotic treatment failure. The pheromone-responsive pCF10 plasmid in Enterococcus faecalis (E. faecalis) mediates antibiotic resistance gene dissemination, but its role in bacterial persister formation remains unclear. This study systematically investigated the regulatory role of pheromone cCF10 in the persister phenotype of pCF10-carrying E. faecalis and its underlying molecular mechanisms. We confirmed that cCF10 enhanced persistence against levofloxacin in OG1RF (pCF10), with the persister frequency increasing from 0.291% to 16.466% upon treatment. Transcriptomic analysis revealed that cCF10 activated the (p)ppGpp-mediated stringent response and downregulated the expression of genes associated with energy-intensive pathways, including those involved in DNA repair, protein folding, and respiration. Concurrently, cCF10 enhanced the expression of genes related to biofilm formation and cell lysis resistance and downregulated components of its own sensing and uptake systems. These findings demonstrate that cCF10 induces transcriptional reprogramming associated with increased persister formation in E. faecalis carrying the pCF10 plasmid and identify potential targets within the stringent response and associated metabolic pathways for the development of anti-persister strategies.