title: H-NS-mediated repression of CRISPR-based immunity in Escherichia coli K12 can be relieved by the transcription activator LeuO.
abstract: The recently discovered prokaryotic CRISPR/Cas defence system provides immunity against viral infections and plasmid conjugation. It has been demonstrated that in Escherichia coli transcription of the Cascade genes (casABCDE) and to some extent the CRISPR array is repressed by heat-stable nucleoid-structuring (H-NS) protein, a global transcriptional repressor. Here we elaborate on the control of the E. coli CRISPR/Cas system, and study the effect on CRISPR-based anti-viral immunity. Transformation of wild-type E. coli K12 with CRISPR spacers that are complementary to phage Lambda does not lead to detectable protection against Lambda infection. However, when an H-NS mutant of E. coli K12 is transformed with the same anti-Lambda CRISPR, this does result in reduced sensitivity to phage infection. In addition, it is demonstrated that LeuO, a LysR-type transcription factor, binds to two sites flanking the casA promoter and the H-NS nucleation site, resulting in derepression of casABCDE12 transcription. Overexpression of LeuO in E. coli K12 containing an anti-Lambda CRISPR leads to an enhanced protection against phage infection. This study demonstrates that in E. coli H-NS and LeuO are antagonistic regulators of CRISPR-based immunity.
authors.affiliation: Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, the Netherlands.
authors.forename: Edze R
authors.initials: ER
authors.lastname: Westra
authors.forename: Umit
authors.initials: U
authors.lastname: Pul
authors.forename: Nadja
authors.initials: N
authors.lastname: Heidrich
authors.forename: Matthijs M
authors.initials: MM
authors.lastname: Jore
authors.forename: Magnus
authors.initials: M
authors.lastname: Lundgren
authors.forename: Thomas
authors.initials: T
authors.lastname: Stratmann
authors.forename: Reinhild
authors.initials: R
authors.lastname: Wurm
authors.forename: Amanda
authors.initials: A
authors.lastname: Raine
authors.forename: Melina
authors.initials: M
authors.lastname: Mescher
authors.forename: Luc
authors.initials: L
authors.lastname: Van Heereveld
authors.forename: Marieke
authors.initials: M
authors.lastname: Mastop
authors.forename: E Gerhart H
authors.initials: EG
authors.lastname: Wagner
authors.forename: Karin
authors.initials: K
authors.lastname: Schnetz
authors.forename: John
authors.initials: J
authors.lastname: Van Der Oost
authors.forename: Rolf
authors.initials: R
authors.lastname: Wagner
authors.forename: Stan J J
authors.initials: SJ
authors.lastname: Brouns
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D010582:Bacteriophage lambda / Q000502:physiology; D001483:Base Sequence; D003001:Cloning, Molecular; D018983:DNA Footprinting; D004269:DNA, Bacterial / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism*; D048168:Escherichia coli K12 / Q000235:genetics* / Q000276:immunology* / Q000821:virology; D029968:Escherichia coli Proteins / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D021901:DNA, Intergenic; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C031908:H-NS protein, bacteria; C430340:LeuO protein, E coli; D014157:Transcription Factors