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Function of the CRISPR-Cas System of the Human Pathogen Clostridium difficile.

Boudry P, Semenova E, Monot M, Datsenko KA, Lopatina A, Sekulovic O, Ospina-Bedoya M, Fortier LC, Severinov K, Dupuy B, Soutourina O - MBio (2015)

Bottom Line: Clostridium difficile is the major cause of nosocomial infections associated with antibiotic therapy worldwide.We provide experimental evidence for the function of the C. difficile CRISPR system against plasmid DNA and bacteriophages.These data demonstrate the original features of active C. difficile CRISPR system and bring important insights into the interactions of this major enteropathogen with foreign DNA invaders during its infection cycle.

View Article: PubMed Central - PubMed

Affiliation: Laboratoire Pathogenèse des Bactéries Anaérobies, Institut Pasteur, Paris, France Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France.

No MeSH data available.


Related in: MedlinePlus

Positions of CRISPR-Cas I-B loci in C. difficile strains 630 and R20291. (A) Schematic view of the genomic locations of expressed CRISPR arrays in strains 630 and R20291. CRISPR arrays (CR) are numbered according to the CRISPRdb database. Arrowheads indicate the array position, and transcriptional orientation is indicated by colors as follows: green for the plus or coding strand and blue for the minus or noncoding strand. The locations of associated cas operons, prophage regions, and replication origin (ori) are indicated. The right and left replichores are shown by black arrowheads. (B) Organization of the operons for the complete (CD2982-CD2975) and partial cas operon (CD2455-CD2451) from C. difficile strain 630 (left) and for the complete (CDR20291_2817-2810) and partial cas operons (CDR20291_2348-2344 and CDR20291_2998-2994) from C. difficile strain R20291 (right). The same color was used for homologous cas genes.
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fig1: Positions of CRISPR-Cas I-B loci in C. difficile strains 630 and R20291. (A) Schematic view of the genomic locations of expressed CRISPR arrays in strains 630 and R20291. CRISPR arrays (CR) are numbered according to the CRISPRdb database. Arrowheads indicate the array position, and transcriptional orientation is indicated by colors as follows: green for the plus or coding strand and blue for the minus or noncoding strand. The locations of associated cas operons, prophage regions, and replication origin (ori) are indicated. The right and left replichores are shown by black arrowheads. (B) Organization of the operons for the complete (CD2982-CD2975) and partial cas operon (CD2455-CD2451) from C. difficile strain 630 (left) and for the complete (CDR20291_2817-2810) and partial cas operons (CDR20291_2348-2344 and CDR20291_2998-2994) from C. difficile strain R20291 (right). The same color was used for homologous cas genes.

Mentions: Comparative analysis of subtype I-B CRISPR-Cas systems in the C. difficile reference strain 630 and the PCR ribotype 027 epidemic strain R20291 was performed. According to the CRISPRdb database, C. difficile strain 630 carries genes that encode 12 potential CRISPR arrays, 5 of which are located within prophage regions (Fig. 1; see Table S1 in the supplemental material) (11). For further reference, we used the CRISPRdb numbering for the C. difficile strain 630 CRISPR arrays and numbered spacers within each array according to the identified transcriptional order. All 12 arrays are actively expressed and processed into crRNA under laboratory conditions (24) (see Fig. S1 in the supplemental material). The distribution of CRISPR arrays throughout the chromosome and their orientation appear to be nonrandom (Fig. 1A). CRISPR 3/4, 6, 7, 8, and 9 are transcribed from one DNA strand in the clockwise direction, and CRISPR 10, 11, 12, 15/16, 17 are transcribed from a different strand in the counterclockwise direction. The cas gene cluster CD2982-CD2975 encoding Cas1 to Cas8 proteins is located near the CRISPR 17 array. An additional incomplete cas operon CD2455-CD2451 lacking the universal cas1 component as well as the cas2 and cas4 genes (Fig. 1B) is found close to the CRISPR 12 array (Fig. 1A). Both cas operons are transcribed according to our RNA-seq data.


Function of the CRISPR-Cas System of the Human Pathogen Clostridium difficile.

Boudry P, Semenova E, Monot M, Datsenko KA, Lopatina A, Sekulovic O, Ospina-Bedoya M, Fortier LC, Severinov K, Dupuy B, Soutourina O - MBio (2015)

Positions of CRISPR-Cas I-B loci in C. difficile strains 630 and R20291. (A) Schematic view of the genomic locations of expressed CRISPR arrays in strains 630 and R20291. CRISPR arrays (CR) are numbered according to the CRISPRdb database. Arrowheads indicate the array position, and transcriptional orientation is indicated by colors as follows: green for the plus or coding strand and blue for the minus or noncoding strand. The locations of associated cas operons, prophage regions, and replication origin (ori) are indicated. The right and left replichores are shown by black arrowheads. (B) Organization of the operons for the complete (CD2982-CD2975) and partial cas operon (CD2455-CD2451) from C. difficile strain 630 (left) and for the complete (CDR20291_2817-2810) and partial cas operons (CDR20291_2348-2344 and CDR20291_2998-2994) from C. difficile strain R20291 (right). The same color was used for homologous cas genes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556805&req=5

fig1: Positions of CRISPR-Cas I-B loci in C. difficile strains 630 and R20291. (A) Schematic view of the genomic locations of expressed CRISPR arrays in strains 630 and R20291. CRISPR arrays (CR) are numbered according to the CRISPRdb database. Arrowheads indicate the array position, and transcriptional orientation is indicated by colors as follows: green for the plus or coding strand and blue for the minus or noncoding strand. The locations of associated cas operons, prophage regions, and replication origin (ori) are indicated. The right and left replichores are shown by black arrowheads. (B) Organization of the operons for the complete (CD2982-CD2975) and partial cas operon (CD2455-CD2451) from C. difficile strain 630 (left) and for the complete (CDR20291_2817-2810) and partial cas operons (CDR20291_2348-2344 and CDR20291_2998-2994) from C. difficile strain R20291 (right). The same color was used for homologous cas genes.
Mentions: Comparative analysis of subtype I-B CRISPR-Cas systems in the C. difficile reference strain 630 and the PCR ribotype 027 epidemic strain R20291 was performed. According to the CRISPRdb database, C. difficile strain 630 carries genes that encode 12 potential CRISPR arrays, 5 of which are located within prophage regions (Fig. 1; see Table S1 in the supplemental material) (11). For further reference, we used the CRISPRdb numbering for the C. difficile strain 630 CRISPR arrays and numbered spacers within each array according to the identified transcriptional order. All 12 arrays are actively expressed and processed into crRNA under laboratory conditions (24) (see Fig. S1 in the supplemental material). The distribution of CRISPR arrays throughout the chromosome and their orientation appear to be nonrandom (Fig. 1A). CRISPR 3/4, 6, 7, 8, and 9 are transcribed from one DNA strand in the clockwise direction, and CRISPR 10, 11, 12, 15/16, 17 are transcribed from a different strand in the counterclockwise direction. The cas gene cluster CD2982-CD2975 encoding Cas1 to Cas8 proteins is located near the CRISPR 17 array. An additional incomplete cas operon CD2455-CD2451 lacking the universal cas1 component as well as the cas2 and cas4 genes (Fig. 1B) is found close to the CRISPR 12 array (Fig. 1A). Both cas operons are transcribed according to our RNA-seq data.

Bottom Line: Clostridium difficile is the major cause of nosocomial infections associated with antibiotic therapy worldwide.We provide experimental evidence for the function of the C. difficile CRISPR system against plasmid DNA and bacteriophages.These data demonstrate the original features of active C. difficile CRISPR system and bring important insights into the interactions of this major enteropathogen with foreign DNA invaders during its infection cycle.

View Article: PubMed Central - PubMed

Affiliation: Laboratoire Pathogenèse des Bactéries Anaérobies, Institut Pasteur, Paris, France Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France.

No MeSH data available.


Related in: MedlinePlus