Function of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>

ABSTRACT Clostridium difficile is the cause of most frequently occurring nosocomial diarrhea worldwide. As an enteropathogen, C. difficile must be exposed to multiple exogenous genetic elements in bacteriophage-rich gut communities. CRISPR (clustered regularly interspaced short palindromic repeats)-...

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Autores principales:	Pierre Boudry, Ekaterina Semenova, Marc Monot, Kirill A. Datsenko, Anna Lopatina, Ognjen Sekulovic, Maicol Ospina-Bedoya, Louis-Charles Fortier, Konstantin Severinov, Bruno Dupuy, Olga Soutourina
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2015
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/782d8722c26d456a996ee365d97ca4f4
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spelling	oai:doaj.org-article:782d8722c26d456a996ee365d97ca4f42021-11-15T15:41:30ZFunction of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>10.1128/mBio.01112-152150-7511https://doaj.org/article/782d8722c26d456a996ee365d97ca4f42015-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01112-15https://doaj.org/toc/2150-7511ABSTRACT Clostridium difficile is the cause of most frequently occurring nosocomial diarrhea worldwide. As an enteropathogen, C. difficile must be exposed to multiple exogenous genetic elements in bacteriophage-rich gut communities. CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems allow bacteria to adapt to foreign genetic invaders. Our recent data revealed active expression and processing of CRISPR RNAs from multiple type I-B CRISPR arrays in C. difficile reference strain 630. Here, we demonstrate active expression of CRISPR arrays in strain R20291, an epidemic C. difficile strain. Through genome sequencing and host range analysis of several new C. difficile phages and plasmid conjugation experiments, we provide evidence of defensive function of the CRISPR-Cas system in both C. difficile strains. We further demonstrate that C. difficile Cas proteins are capable of interference in a heterologous host, Escherichia coli. These data set the stage for mechanistic and physiological analyses of CRISPR-Cas-mediated interactions of important global human pathogen with its genetic parasites. IMPORTANCE Clostridium difficile is the major cause of nosocomial infections associated with antibiotic therapy worldwide. To survive in bacteriophage-rich gut communities, enteropathogens must develop efficient systems for defense against foreign DNA elements. CRISPR-Cas systems have recently taken center stage among various anti-invader bacterial defense systems. 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.Pierre BoudryEkaterina SemenovaMarc MonotKirill A. DatsenkoAnna LopatinaOgnjen SekulovicMaicol Ospina-BedoyaLouis-Charles FortierKonstantin SeverinovBruno DupuyOlga SoutourinaAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 6, Iss 5 (2015)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Pierre Boudry Ekaterina Semenova Marc Monot Kirill A. Datsenko Anna Lopatina Ognjen Sekulovic Maicol Ospina-Bedoya Louis-Charles Fortier Konstantin Severinov Bruno Dupuy Olga Soutourina Function of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>
description	ABSTRACT Clostridium difficile is the cause of most frequently occurring nosocomial diarrhea worldwide. As an enteropathogen, C. difficile must be exposed to multiple exogenous genetic elements in bacteriophage-rich gut communities. CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems allow bacteria to adapt to foreign genetic invaders. Our recent data revealed active expression and processing of CRISPR RNAs from multiple type I-B CRISPR arrays in C. difficile reference strain 630. Here, we demonstrate active expression of CRISPR arrays in strain R20291, an epidemic C. difficile strain. Through genome sequencing and host range analysis of several new C. difficile phages and plasmid conjugation experiments, we provide evidence of defensive function of the CRISPR-Cas system in both C. difficile strains. We further demonstrate that C. difficile Cas proteins are capable of interference in a heterologous host, Escherichia coli. These data set the stage for mechanistic and physiological analyses of CRISPR-Cas-mediated interactions of important global human pathogen with its genetic parasites. IMPORTANCE Clostridium difficile is the major cause of nosocomial infections associated with antibiotic therapy worldwide. To survive in bacteriophage-rich gut communities, enteropathogens must develop efficient systems for defense against foreign DNA elements. CRISPR-Cas systems have recently taken center stage among various anti-invader bacterial defense systems. 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.
format	article
author	Pierre Boudry Ekaterina Semenova Marc Monot Kirill A. Datsenko Anna Lopatina Ognjen Sekulovic Maicol Ospina-Bedoya Louis-Charles Fortier Konstantin Severinov Bruno Dupuy Olga Soutourina
author_facet	Pierre Boudry Ekaterina Semenova Marc Monot Kirill A. Datsenko Anna Lopatina Ognjen Sekulovic Maicol Ospina-Bedoya Louis-Charles Fortier Konstantin Severinov Bruno Dupuy Olga Soutourina
author_sort	Pierre Boudry
title	Function of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>
title_short	Function of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>
title_full	Function of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>
title_fullStr	Function of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>
title_full_unstemmed	Function of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>
title_sort	function of the crispr-cas system of the human pathogen <named-content content-type="genus-species">clostridium difficile</named-content>
publisher	American Society for Microbiology
publishDate	2015
url	https://doaj.org/article/782d8722c26d456a996ee365d97ca4f4
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Function of the CRISPR-Cas System of the Human Pathogen <named-content content-type="genus-species">Clostridium difficile</named-content>

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