Transcriptional control of two distinct lactococcal plasmid-encoded conjugation systems

•Active promoters in pNP40 and pUC11B conjugation clusters are identified.•Regulatory elements of said clusters include the conjugation-associated relaxases.•Relaxase expression of both systems appears to be autoregulated.•Tra20 may regulate pNP40 conjugation in an uncharacterized manner. Lactococcal conjugative plasmids are poorly characterized compared to those harbored by numerous other Gram-positive bacteria, despite their significance in dairy fermentations and starter culture development. Furthermore, the transcriptional landscape of these lactococcal conjugation systems and their regulation have not been studied in any detail. Lactococcal plasmids pNP40 and pUC11B possess two genetically distinct and prevalent conjugation systems. Here, we describe the detailed transcriptional analysis of the pNP40 and pUC11B conjugation-associated gene clusters, revealing three and five promoters, respectively, for which the corresponding transcriptional start sites were identified. Regulation of several of these promoters, and therefore conjugation, is shown to involve the individual or concerted activities of the corresponding relaxase and transcriptional repressor(s) encoded by each conjugative plasmid. This work highlights how the conjugative potential of these systems may be unlocked, with significant implications for the starter culture and food fermentation industry. [Display omitted]