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High pressure activation of the Mrr restriction endonuclease in Escherichia coli involves tetramer dissociation
Rensselaer Polytech Inst, Dept Biol Sci, Troy, NY 12180 USA.;Univ Montpellier, CNRS UMR5048, INSERM U1054, Ctr Biochim Struct, F-34000 Montpellier, France..
Katholieke Univ Leuven, Food Microbiol Lab, Dept Microbial & Mol Syst, B-3001 Leuven, Belgium..
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Microbiology. Katholieke Univ Leuven, Food Microbiol Lab, Dept Microbial & Mol Syst, B-3001 Leuven, Belgium.
Katholieke Univ Leuven, Food Microbiol Lab, Dept Microbial & Mol Syst, B-3001 Leuven, Belgium..
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2017 (English)In: Nucleic Acids Research, ISSN 0305-1048, E-ISSN 1362-4962, Vol. 45, no 9, 5323-5332 p.Article in journal (Refereed) Published
Abstract [en]

A sub-lethal hydrostatic pressure (HP) shock of similar to 100 MPa elicits a RecA-dependent DNA damage (SOS) response in Escherichia coli K-12, despite the fact that pressure cannot compromise the covalent integrity of DNA. Prior screens for HP resistance identified Mrr (Methylated adenine Recognition and Restriction), a Type IV restriction endonuclease (REase), as instigator for this enigmatic HP-induced SOS response. Type IV REases tend to target modified DNA sites, and E. coli Mrr activity was previously shown to be elicited by expression of the foreign M. HhaII Type II methytransferase (MTase), as well. Here we measured the concentration and stoichiometry of a functional GFP-Mrr fusion protein using in vivo fluorescence fluctuation microscopy. Our results demonstrate that Mrr is a tetramer in unstressed cells, but shifts to a dimer after HP shock or co-expression with M. HhaII. Based on the differences in reversibility of tetramer dissociation observed for wild-type GFP-Mrr and a catalytic mutant upon HP shock compared to M. HhaII expression, we propose a model by which (i) HP triggers Mrr activity by directly pushing inactive Mrr tetramers to dissociate into active Mrr dimers, while (ii) M. HhaII triggers Mrr activity by creating high affinity target sites on the chromosome, which pull the equilibrium from inactive tetrameric Mrr toward active dimer.

Place, publisher, year, edition, pages
OXFORD UNIV PRESS , 2017. Vol. 45, no 9, 5323-5332 p.
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Cell and Molecular Biology
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URN: urn:nbn:se:uu:diva-326240DOI: 10.1093/nar/gkx192ISI: 000402064200035PubMedID: 28369499OAI: oai:DiVA.org:uu-326240DiVA: diva2:1120405
Available from: 2017-07-06 Created: 2017-07-06 Last updated: 2017-07-06Bibliographically approved

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