title: Functional Analysis of Porphyromonas gingivalis W83 CRISPR-Cas Systems.
abstract: The CRISPR-Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated genes) system provides prokaryotic cells with an adaptive and heritable immune response to foreign genetic elements, such as viruses, plasmids, and transposons. It is present in the majority of Archaea and almost half of species of Bacteria. Porphyromonas gingivalis is an important human pathogen that has been proven to be an etiological agent of periodontitis and has been linked to systemic conditions, such as rheumatoid arthritis and cardiovascular disease. At least 95% of clinical strains of P. gingivalis carry CRISPR arrays, suggesting that these arrays play an important function in vivo. Here we show that all four CRISPR arrays present in the P. gingivalis W83 genome are transcribed. For one of the arrays, we demonstrate in vivo activity against double-stranded DNA constructs containing protospacer sequences accompanied at the 3' end by an NGG protospacer-adjacent motif (PAM). Most of the 44 spacers present in the genome of P. gingivalis W83 share no significant similarity with any known sequences, although 4 spacers are similar to sequences from bacteria found in the oral cavity and the gastrointestinal tract. Four spacers match genomic sequences of the host; however, none of these is flanked at its 3' terminus by the appropriate PAM element.
  
  
  OBJECTIVE
  The CRISPR-Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated genes) system is a unique system that provides prokaryotic cells with an adaptive and heritable immunity. In this report, we show that the CRISPR-Cas system of P. gingivalis, an important human pathogen associated with periodontitis and possibly also other conditions, such as rheumatoid arthritis and cardiovascular disease, is active and provides protection from foreign genetic elements. Importantly, the data presented here may be useful for better understanding the communication between cells in larger bacterial communities and, consequently, the process of disease development and progression.
authors.affiliation: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland.
authors.forename: Michał
authors.initials: M
authors.lastname: Burmistrz
authors.affiliation: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland.
authors.forename: Bartosz
authors.initials: B
authors.lastname: Dudek
authors.affiliation: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland.
authors.forename: Dominika
authors.initials: D
authors.lastname: Staniec
authors.affiliation: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland.
authors.forename: Jose Ignacio
authors.initials: JI
authors.lastname: Rodriguez Martinez
authors.affiliation: Laboratory of Structural Biology, International Institute of Molecular and Cell Biology, Warsaw, Poland Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: Matthias
authors.initials: M
authors.lastname: Bochtler
authors.affiliation: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland Oral Health and Systemic Disease Research Group, School of Dentistry, University of Louisville, Louisville, Kentucky, USA.
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
authors.affiliation: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, Poland Malopolska Center of Biotechnology, Jagiellonian University, Cracow, Poland k.a.pyrc@uj.edu.pl.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Pyrc
mesh_terms: D000595:Amino Acid Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics; D003470:Culture Media / Q000737:chemistry; D004247:DNA / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D056426:Genetic Loci; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D016966:Porphyromonas gingivalis / Q000235:genetics*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D015533:Transcriptional Activation
chemical_list: D003470:Culture Media; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004247:DNA