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Macrolide resistance, emerging in  Streptococcus pneumoniae  and other Gram-positive bacteria, is increasingly due to efflux pumps encoded by  mef/mel(msr)  operons found on discrete mobile genetic elements. The regulation of  mef/mel(msr)  in these elements is not well understood. We identified the  mef(E)/mel  transcriptional start, localized the  mef(E)/mel  promoter, and demonstrated attenuation of transcription as a mechanism of regulation of macrolide-inducible  mef -mediated macrolide resistance in  S .  pneumoniae . The  mef(E)/mel  transcriptional start site was a guanine 327 bp upstream of  mef(E) . Consensus pneumococcal promoter -10 (5′-TATACT-3′) and -35 (5′-TTGAAC-3′) boxes separated by 17 bp were identified 7 bp upstream of the start site. Analysis of the predicted secondary structure of the 327 5’ region identified four pairs of inverted repeats R1-R8 predicted to fold into stem-loops, a small leader peptide [MTASMRLR, (Mef(E)L)] required for macrolide induction and a Rho-independent transcription terminator. RNA-seq analyses provided confirmation of transcriptional attenuation. In addition, expression of  mef(E)L  was also influenced by  mef(E)L -dependent mRNA stability. The regulatory region 5’ of  mef(E)  was highly conserved in other  mef/mel(msr) -containing elements including  Tn 1207.1 and the 5612IQ complex in pneumococci and  Tn 1207.3 in Group A streptococci, indicating a regulatory mechanism common to a wide variety of Gram-positive bacteria containing  mef/mel(msr)  elements.

plos one

Transcriptional Attenuation Controls Macrolide Inducible Efflux and Resistance in Streptococcus pneumoniae and in Other Gram-Positive Bacteria Containing mef/mel(msr(D)) Elements