Analysis of sigma(54)-dependent genes in Enterococcus faecalis: a mannose PTS permease (EII(Man)) is involved in sensitivity to a bacteriocin, mesentericin Y105.
The sigma(54) RNA polymerase subunit has a prominent role in susceptibility of Listeria monocytogenes and Enterococcus faecalis to mesentericin Y105, a class IIa bacteriocin. Consequently, sigma(54)-dependent genes as well as specific activators also required for expression of these genes were sought. Five putative sigma(54)-associated activators were detected in the genome of E. faecalis V583, and all but one could activate the transcription of permease genes belonging to sugar phosphotransferase systems (PTSs). Interestingly, these activators display a helicase signature not yet reported in this activator family, which could explain the ATP-dependent mechanism of DNA unwinding preceding the start of transcription. To find which activator is linked to susceptibility of E. faecalis to mesentericin Y105, their respective genes were subsequently interrupted. Among them, only mptR gene interruption led to a resistance phenotype. Immediately downstream from mptR, a putative sigma(54)-dependent operon was found to encode a mannose PTS permease, namely EII(t)(Man). Moreover, in liquid culture, glucose and mannose induced the sensitivity of E. faecalis to mesentericin Y105. Since sugars have previously been reported to induce PTS permease expression, it appears that EII(t)(Man) expression, presumably induced in the presence of glucose and mannose, leads to an enhanced sensitivity of E. faecalis to the bacteriocin. Additional information was gained from knockouts within the permease operon. Interruption of the distal mptD gene, which encodes the IID subunit of EII(t)(Man), strikingly led to resistance to mesentericin Y105. Moreover, MptD appears to be a peculiar membrane subunit, bearing an additional domain compared to most known IID subunits. According to these results, EII(t)(Man) is clearly involved in susceptibility to mesentericin Y105 and could even be its receptor at the E. faecalis surface. Finally, it is hypothesized that MptD could be responsible for the targeting specificity, via an interaction between its additional domain and mesentericin Y105.
Héchard
Yann
Y
Laboratoire de Microbiologie Fondamentale et Appliquée, CNRS FRE 2224, IBMIG, UFR Sciences, 40 avenue du Recteur Pineau, 86022 Poitiers Cedex, France1.
Pelletier
Christelle
C
Laboratoire de Microbiologie Fondamentale et Appliquée, CNRS FRE 2224, IBMIG, UFR Sciences, 40 avenue du Recteur Pineau, 86022 Poitiers Cedex, France1.
Cenatiempo
Yves
Y
Laboratoire de Microbiologie Fondamentale et Appliquée, CNRS FRE 2224, IBMIG, UFR Sciences, 40 avenue du Recteur Pineau, 86022 Poitiers Cedex, France1.
Frère
Jacques
J
Laboratoire de Microbiologie Fondamentale et Appliquée, CNRS FRE 2224, IBMIG, UFR Sciences, 40 avenue du Recteur Pineau, 86022 Poitiers Cedex, France1.
D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000378:metabolism / Q000494:pharmacology; D003470:Culture Media; D004268:DNA-Binding Proteins; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000378:metabolism; D013293:Enterococcus faecalis / Q000235:genetics* / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D008358:Mannose / Q000378:metabolism*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009876:Operon; D010641:Phenotype; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics* / Q000378:metabolism; D051877:RNA Polymerase Sigma 54; D016415:Sequence Alignment; D012808:Sigma Factor / Q000235:genetics* / Q000378:metabolism
D001430:Bacteriocins; D003470:Culture Media; D004268:DNA-Binding Proteins; D012808:Sigma Factor; C079275:mesentericin Y105 protein, Leuconostoc mesenteroides; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C037992:phosphoenolpyruvate-mannose phosphotransferase; D012321:DNA-Directed RNA Polymerases; D051877:RNA Polymerase Sigma 54; D005947:Glucose; D008358:Mannose