title: Generation of a CRISPR database for Yersinia pseudotuberculosis complex and role of CRISPR-based immunity in conjugation.
abstract: The clustered regularly interspaced short palindromic repeat - CRISPR-associated genes (CRISPR-Cas) system is used by bacteria and archaea against invading conjugative plasmids or bacteriophages. Central to this immunity system are genomic CRISPR loci that contain fragments of invading DNA. These are maintained as spacers in the CRISPR loci between direct repeats and the spacer composition in any bacterium reflects its evolutionary history. We analysed the CRISPR locus sequences of 335 Yersinia pseudotuberculosis complex strains. Altogether 1902 different spacer sequences were identified and these were used to generate a database for the spacer sequences. Only ∼10% of the spacer sequences found matching sequences. In addition, surprisingly few spacers were shared by Yersinia pestis and Y. pseudotuberculosis strains. Interestingly, 32 different protospacers were present in the conjugative plasmid pYptb32953. The corresponding spacers were identified from 35 different Y. pseudotuberculosis strains indicating that these strains had encountered pYptb32953 earlier. In conjugation experiments, pYptb32953-specific spacers generally prevented conjugation with spacer-positive and spacer-free strains. However, some strains with one to four spacers were invaded by pYptb32953 and some spacer-free strains were fully resistant. Also some spacer-positive strains were intermediate resistant to conjugation. This suggests that one or more other defence systems are determining conjugation efficiency independent of the CRISPR-Cas system.
authors.affiliation: Research and Development Department, Centre for Military Medicine, Helsinki, Finland.
authors.forename: Katja A
authors.initials: KA
authors.lastname: Koskela
authors.affiliation: Department of Bacteriology and Immunology, Haartman Institute and Research Programs Unit, Immunobiology, University of Helsinki, PO Box 21, 00014, Helsinki, Finland.
authors.forename: Laura
authors.initials: L
authors.lastname: Mattinen
authors.affiliation: Research and Development Department, Centre for Military Medicine, Helsinki, Finland.|Department of Bacteriology and Immunology, Haartman Institute and Research Programs Unit, Immunobiology, University of Helsinki, PO Box 21, 00014, Helsinki, Finland.
authors.forename: Laura
authors.initials: L
authors.lastname: Kalin-Mänttäri
authors.affiliation: Univ Paris-Sud, Institut de Génétique et Microbiologie, UMR8621, Orsay, France.|CNRS, Orsay, France.|ENSTA ParisTech, Palaiseau, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Vergnaud
authors.affiliation: Univ Paris-Sud, Institut de Génétique et Microbiologie, UMR8621, Orsay, France.|CNRS, Orsay, France.|DGA/MNRBC, Vert le Petit, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Gorgé
authors.affiliation: Research and Development Department, Centre for Military Medicine, Helsinki, Finland.
authors.forename: Simo
authors.initials: S
authors.lastname: Nikkari
authors.affiliation: Department of Bacteriology and Immunology, Haartman Institute and Research Programs Unit, Immunobiology, University of Helsinki, PO Box 21, 00014, Helsinki, Finland.|Helsinki University Central Hospital Laboratory Diagnostics, Helsinki, Finland.
authors.forename: Mikael
authors.initials: M
authors.lastname: Skurnik
mesh_terms: D001435:Bacteriophages / Q000276:immunology; D001483:Base Sequence; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics / Q000276:immunology*; D003227:Conjugation, Genetic / Q000276:immunology*; D030561:Databases, Nucleic Acid; D023281:Genomics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000276:immunology*; D015010:Yersinia pestis / Q000235:genetics; D015011:Yersinia pseudotuberculosis / Q000145:classification / Q000235:genetics*