H-NS mediated repression of CRISPR-based immunity in Escherichia coli K12 can be relieved by the transcription activator LeuO

• Published in: Molecular Microbiology
• Language: English
• Published: Wiley-Blackwell 2011
• Subjects: Life Sciences; Lambda; Heine; issoleie
• Online Access: http://hdl.handle.net/2262/49752; https://doi.org/10.1111/j.1365-2958.2010.07315.x

Abstract The recently discovered prokaryotic CRISPR/Cas defense 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. Over-expression 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. : Pul, Umit, [e-mail] merged with this user on 21-Jun-2010 by Helmann, John (Pul, Umit) : Heidrich, Nadja, [e-mail] merged with this user on 15-Jun-2010 by Helmann, John (Heidrich, Nadja) : Lundgren, Magnus, [e-mail] merged with this user on 17-Jun-2010 by Helmann, John (Lundgren, Magnus) : imported for journal id 147 on Jul 20, 2001, no letter sent29 Nov 2001 account with Record No. 258003 merged into this account by Ms. Wendy Priest (Wagner, E. Gerhart H.) : 11 Dec 2001 account with Record No. 257218 merged into this account by Prof. Chris Higgins (Schnetz, Karin) : 19 Feb 2004 - Added using 'Add Referee on the fly' by Prof. Tony Pugsley (Wagner, Rolf) [e-mail] (Westra, Edze Rients) [e-mail] (Pul, Umit) [e-mail] (Heidrich, Nadja) [e-mail] (Jore, Matthijs Miklas) [e-mail] (Lundgren, Magnus) [e-mail] (Stratmann, Thomas) [e-mail] (Wurm, Reinhild) [e-mail] (Raine, Amanda) [e-mail] (Mescher, Melina) [e-mail] (Heereveld, Luc van) [e-mail] (Mastop, Marieke) [e-mail] (Wagner, E. Gerhart H.) [e-mail] (Schnetz, Karin) [e-mail] (Van der Oost, John) [e-mail] (Wagner, Rolf) [e-mail] (Brouns, Stan J.J.) Wageningen University, Microbiology - Drijenplein 10--> - 6703 HB - Wageningen - NETHERLANDS (Westra, Edze Rients) Heinrich-Heine-Universitat Dusseldorf, Dusseldorf, Germany, Molecular Biology of Prokaryotes - Dusseldorf - GERMANY (Pul, Umit) Uppsala University, Institute of Cell and Molecular Biology - Uppsala - SWEDEN (Heidrich, Nadja) Wageningen University, Laboratory of Microbiology - Wageningen - NETHERLANDS (Jore, Matthijs Miklas) Uppsala University, Institute of Cell and Molecular Biology - Uppsala - SWEDEN (Lundgren, Magnus) University of Cologne, Institute for Genetics - Cologne - GERMANY (Stratmann, Thomas) Heinrich-Heine-Universitat Dusseldorf, Molecular Biology of Prokaryotes - Dusseldorf - GERMANY (Wurm, Reinhild) Uppdala University, Department of Cell and Molecular Biology - Uppsala - SWEDEN (Raine, Amanda) Heinrich-Heine-Universitat Dusseldorf, Molecular Biology of Prokaryotes - Dusseldorf - GERMANY (Mescher, Melina) Wageningen University, Laboratory of Microbiology - Wageningen - NETHERLANDS (Heereveld, Luc van) Wageningen University, Laboratory of Microbiology - Wageningen - NETHERLANDS (Mastop, Marieke) Uppsala university, Department of Cell and Molecular Biology - Biomedical Center--> , Box 596--> - S-751 24 - Uppsala - SWEDEN (Wagner, E. Gerhart H.) SWEDEN (Wagner, E. Gerhart H.) University of Cologne, Institute for Genetics - Zuelpicher Str. 47--> - 50674 - Cologne - GERMANY (Schnetz, Karin) GERMANY (Schnetz, Karin) Wageningen University, Laboratory of Microbiology - Wageningen - NETHERLANDS (Van der Oost, John) Heinrich-Heine-Universitat Dusseldorf, Molecular Biology of Prokaryotes - Dusseldorf - GERMANY (Wagner, Rolf) Wageningen University, Laboratory of Microbiology - Wageningen - NETHERLANDS (Brouns, Stan J.J.) 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