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This Article Full Text Full Text (PDF) Supplementary data All Versions of this Article: mic.0.027508-0v1 155/6/1966    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Google Scholar Articles by van der Ploeg, J. R. PubMed PubMed Citation Articles by van der Ploeg, J. R. Agricola Articles by van der Ploeg, J. R.

Analysis of CRISPR in Streptococcus mutans suggests frequent occurrence of acquired immunity against infection by M102-like bacteriophages

Institute of Oral Biology, University of Zurich, Plattenstrasse 11, CH-8032 Zurich, Switzerland

Clustered regularly interspaced short palindromic repeats (CRISPR) consist of highly conserved direct repeats interspersed with variable spacer sequences. They can protect bacteria against invasion by foreign DNA elements. The genome sequence of Streptococcus mutans strain UA159 contains two CRISPR loci, designated CRISPR1 and CRISPR2. The aims of this study were to analyse the organization of CRISPR in further S. mutans strains and to investigate the importance of CRISPR in acquired immunity to M102-like phages. The sequences of CRISPR1 and CRISPR2 arrays were determined for 29 S. mutans strains from different persons. More than half of the CRISPR1 spacers and about 35 % of the CRISPR2 spacers showed sequence similarity with the genome sequence of M102, a virulent siphophage specific for S. mutans. Although only a few spacers matched the phage sequence completely, most of the mismatches had no effect on the amino acid sequences of the phage-encoded proteins. The results suggest that S. mutans is often attacked by M102-like bacteriophages, and that its acquisition of novel phage-derived CRISPR sequences goes along with the presence of S. mutans phages in the environment. Analysis of CRISPR1 of M102-resistant mutants of S. mutans OMZ 381 showed that some of them had acquired novel spacers, and the sequences of all but one of these matched the phage M102 genome sequence. This suggests that the acquisition of the spacers contributed to the resistance against phage infection. However, since not all resistant mutants had new spacers, and since the removal of the CRISPR1 array in one of the mutants and in wild-type strains did not lead to loss of resistance to infection by M102, the acquisition of resistance must be based on further elements as well.

Correspondence
Jan R. van der Ploeg
jan.vanderploeg{at}zzmk.uzh.ch

A supplementary figure and six supplementary tables are available with the online version of this paper. The supplementary figure shows a comparison of part of the cas1 gene from Streptococcus mutans, Streptococcus equi subsp. zooepidemicus and Streptococcus pyogenes strains. The supplementary tables list oligonucleotides used in this study, CRISPR1 and CRISPR2 spacer sequences, sequences of the CRISPR1 and CRISPR2 repeats, and similarity of S. mutans CRISPR spacers to sequences other than those of M102.