title: The Cas6e ribonuclease is not required for interference and adaptation by the E. coli type I-E CRISPR-Cas system.
abstract: CRISPR-Cas are small RNA-based adaptive prokaryotic immunity systems protecting cells from foreign DNA or RNA. Type I CRISPR-Cas systems are composed of a multiprotein complex (Cascade) that, when bound to CRISPR RNA (crRNA), can recognize double-stranded DNA targets and recruit the Cas3 nuclease to destroy target-containing DNA. In the Escherichia coli type I-E CRISPR-Cas system, crRNAs are generated upon transcription of CRISPR arrays consisting of multiple palindromic repeats and intervening spacers through the function of Cas6e endoribonuclease, which cleaves at specific positions of repeat sequences of the CRISPR array transcript. Cas6e is also a component of Cascade. Here, we show that when mature unit-sized crRNAs are provided in a Cas6e-independent manner by transcription termination, the CRISPR-Cas system can function without Cas6e. The results should allow facile interrogation of various targets by type I-E CRISPR-Cas system in E. coli using unit-sized crRNAs generated by transcription.
authors.affiliation: Waksman Institute of Microbiology, Rutgers, the State University of New Jersey, Piscataway, NJ 08854, USA.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Semenova
authors.affiliation: Waksman Institute of Microbiology, Rutgers, the State University of New Jersey, Piscataway, NJ 08854, USA.
authors.forename: Konstantin
authors.initials: K
authors.lastname: Kuznedelov
authors.affiliation: Waksman Institute of Microbiology, Rutgers, the State University of New Jersey, Piscataway, NJ 08854, USA Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Kirill A
authors.initials: KA
authors.lastname: Datsenko
authors.affiliation: Waksman Institute of Microbiology, Rutgers, the State University of New Jersey, Piscataway, NJ 08854, USA Laboratoire Pathogenèse des Bactéries Anaérobies, Institut Pasteur, Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.
authors.forename: Pierre M
authors.initials: PM
authors.lastname: Boudry
authors.affiliation: Skolkovo Institute of Science and Technology, Skolkovo 143025, Russia Institute of Molecular Genetics, Russian Academy of Sciences, Moscow 123182, Russia.
authors.forename: Ekaterina E
authors.initials: EE
authors.lastname: Savitskaya
authors.affiliation: Skolkovo Institute of Science and Technology, Skolkovo 143025, Russia Institute of Gene Biology, Russian Academy of Sciences, Moscow 119334, Russia.
authors.forename: Sofia
authors.initials: S
authors.lastname: Medvedeva
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario M5S 3E5, Canada.
authors.forename: Natalia
authors.initials: N
authors.lastname: Beloglazova
authors.affiliation: Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119234 Moscow, Russia Pirogov Russian National Research Medical University, 117997 Moscow, Russia.
authors.forename: Maria
authors.initials: M
authors.lastname: Logacheva
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario M5S 3E5, Canada.
authors.forename: Alexander F
authors.initials: AF
authors.lastname: Yakunin
authors.affiliation: Waksman Institute of Microbiology, Rutgers, the State University of New Jersey, Piscataway, NJ 08854, USA Skolkovo Institute of Science and Technology, Skolkovo 143025, Russia Institute of Molecular Genetics, Russian Academy of Sciences, Moscow 123182, Russia Institute of Gene Biology, Russian Academy of Sciences, Moscow 119334, Russia Peter the Great St. Petersburg Polytechnic University, St. Petersburg 195251, Russia severik@waksman.rutgers.edu.
authors.forename: Konstantin
authors.initials: K
authors.lastname: Severinov
mesh_terms: D001435:Bacteriophages / Q000235:genetics; D064130:CRISPR-Associated Proteins / Q000378:metabolism / Q000502:physiology*; D064113:CRISPR-Cas Systems; D004722:Endoribonucleases / Q000378:metabolism / Q000502:physiology*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D012313:RNA / Q000378:metabolism; D061806:Transcription Termination, Genetic
chemical_list: D064130:CRISPR-Associated Proteins; D012313:RNA; D004722:Endoribonucleases