Binding and cleavage of nicked substrates by site-specific recombinases XerC and XerD

Journal of Molecular Biology
G BlakelyD J Sherratt

Abstract

In Xer site-specific recombination two related recombinases, XerC and XerD, catalyse strand cleavage and rejoining reactions at a site, dif, in order to ensure normal chromosome segregation during cell division in Escherichia coli. We have used nicked suicide substrates to trap reaction intermediates and show that XerC cleaves the top strand efficiently while XerD is less efficient at cleaving the bottom strand of dif. Recombinase-mediated cleavage positions are separated by six base pairs and occur at either end of the dif central region adjacent to the recombinase binding sites. XerC can cleave the top strand of dif inefficiently in the absence of its partner recombinase during a reaction that does not require intermolecular synapsis. The presence of a nick in the bottom strand of dif allows cooperative interactions between two XerC protomers bound to adjacent binding sites, suggesting that a conserved interaction domain is present in both XerC and XerD. Cooperativity between two identical recombinase protomers does not occur on un-nicked linear DNA. Ethylation interference footprinting of two XerD catalytic mutant proteins suggests that the conserved domain II arginine from the integrase family RHRY tetrad may make direct co...Continue Reading

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Citations

Feb 27, 1999·Molecular Microbiology·L NeilsonD J Sherratt
Jun 1, 1997·The EMBO Journal·L K ArciszewskaD J Sherratt
Oct 6, 1997·The EMBO Journal·H S SubramanyaD B Wigley
Jul 12, 2002·The EMBO Journal·Migena BreguSean D Colloms
Sep 15, 1997·Nucleic Acids Research·D Esposito, J J Scocca
Oct 27, 2009·Nucleic Acids Research·James E GrahamLidia K Arciszewska
Mar 5, 2011·Nucleic Acids Research·Ian GraingeDavid J Sherratt
Sep 19, 2002·Protein Science : a Publication of the Protein Society·Daniel J RigdenMark J Jedrzejas
Nov 16, 2010·Journal of Bacteriology·Nadia M DomínguezJoseph P Dillard
Feb 24, 2001·Journal of Bacteriology·S A SciochettiG W Blakely
Sep 4, 2009·PloS One·Christophe Carnoy, Claude-Alain Roten
Jun 11, 1999·Molecular Microbiology·A J Spiers, D J Sherratt
May 13, 2006·Molecular Microbiology·Karolina MalanowskaJeffrey F Gardner
Jun 19, 2016·Proceedings of the National Academy of Sciences of the United States of America·Florian FournesPhilippe Rousseau
Apr 27, 2016·Journal of Molecular Biology·Bryce L Lunt, Graham F Hatfull
Jun 22, 2000·Journal of Molecular Biology·L K ArciszewskaD J Sherratt
Jun 28, 2005·Molecular Microbiology·Sarah M McLeodMatthew K Waldor
May 5, 2004·The Journal of Biological Chemistry·Claire LetzelterMarie-Claude Serre
Nov 22, 2013·Journal of Virology·Michèle CoddevillePaul Ritzenthaler
Sep 25, 2019·Journal of Bacteriology·Ali FarrokhiGeorge Szatmari
Apr 1, 2000·The Journal of Biological Chemistry·G W BlakelyD J Sherratt
Feb 26, 1999·The Journal of Biological Chemistry·I Grainge, D J Sherratt
Nov 4, 2004·Molecular Microbiology·Sarah M McLeod, Matthew K Waldor
Nov 15, 2002·Methods : a Companion to Methods in Enzymology·Kaushik Ghosh, Gregory D Van Duyne
Dec 4, 2002·Journal of Molecular Biology·Michael TekleSimone E Nunes-Düby

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