Bacteriocin (mutacin) production by Streptococcus mutans genome sequence reference strain UA159: elucidation of the antimicrobial repertoire by genetic dissection - PubMed (original) (raw)

Bacteriocin (mutacin) production by Streptococcus mutans genome sequence reference strain UA159: elucidation of the antimicrobial repertoire by genetic dissection

John D F Hale et al. Appl Environ Microbiol. 2005 Nov.

Abstract

Streptococcus mutans UA159, the genome sequence reference strain, exhibits nonlantibiotic mutacin activity. In this study, bioinformatic and mutational analyses were employed to demonstrate that the antimicrobial repertoire of strain UA159 includes mutacin IV (specified by the nlm locus) and a newly identified bacteriocin, mutacin V (encoded by SMU.1914c).

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Figures

FIG. 1.

Allelic replacement, by PCR ligation mutagenesis (13), of genes (nlmAB) encoding the peptides comprising mutacin IV. Other putative bacteriocin-encoding genes investigated in this study (see Fig. 2) were also inactivated using this strategy. For clarity, the loci are not drawn to scale. The PCR primers shown are numbered as follows: 1, NlmABUpF; 2, NlmAUpR; 3, NlmADwF; 4, NlmBUpR; 5, NlmBDwF; 6, NlmABDwR. The filled and unfilled circles refer to EcoRI and PstI restriction sites, respectively. The source of ermAM was pSLER1 (pSL1190 [4] containing ermAM cloned into the NdeI site; laboratory collection) and ermAM was always inserted in the same transcriptional orientation as the gene of interest. The locations of the ErmInv PCR primers used to confirm each mutation are also shown. adhE, putative alcohol-acetaldehyde dehydrogenase; nlmA and nlmB, prepeptides of NlmA* and NlmB*, respectively; SMU.149, putative transposase; SMU.152, hypothetical protein.

FIG. 2.

Genomic organization of the open reading frames (ORFs; filled pentagons) encoding potential nonlantibiotic mutacins investigated in this study. For simplicity, the loci are not drawn to scale. ORFs designated by their GenBank locus tags (e.g., SMU.283) encode hypothetical proteins. Note that SMU.282 does not exist. The translational orientation of each ORF is also indicated. nlmTE, ABC transporter required for export of nonlantibiotic mutacins (10); tif2, translation initiation factor 2; rbfA, ribosome binding factor A; copY, putative transcriptional regulator; copAZ, components of a copper transport system; tnp, putative transposase of IS_Smu1_ (1); cslAB, ABC transport system required for natural transformation (10, 18); comCDE, signal transduction system essential for development of natural competence (8); dedA, putative membrane-associated protein.

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Bacteriocin (mutacin) production by Streptococcus mutans genome sequence reference strain UA159: elucidation of the antimicrobial repertoire by genetic dissection - PubMed (original) (raw)