Site-specific recombinases are enzymes that promote precise rearrangements of DNA sequences. They do this by cutting and rejoining the DNA strands at specific positions within a pair of target sites recognized and bound by the recombinase. One group of these enzymes, the serine recombinases, initiates strand exchange by making double-strand breaks in the DNA of the two sites, in an intermediate built around a catalytic tetramer of recombinase subunits. However, these catalytic steps are only the culmination of a complex pathway that begins when recombinase subunits recognize and bind to their target sites as dimers. To form the tetramer-containing reaction intermediate, two dimer-bound sites are brought together by protein dimer–dimer interactions. During or after this initial synapsis step, the recombinase subunit and tetramer conformations change dramatically by repositioning of component subdomains, bringing about a transformation of the enzyme from an inactive to an active configuration. In natural serine recombinase systems, these steps are subject to elaborate regulatory mechanisms in order to ensure that cleavage and rejoining of DNA strands only happen when and where they should, but we and others have identified recombinase mutants that have lost dependence on this regulation, thus facilitating the study of the basic steps leading to catalysis. We describe how our studies on activated mutants of two serine recombinases, Tn3 resolvase and Sin, are providing us with insights into the structural changes that occur before catalysis of strand exchange, and how these steps in the reaction pathway are regulated.
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April 2011
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Conference Article|
March 22 2011
Intermediates in serine recombinase-mediated site-specific recombination
W. Marshall Stark;
W. Marshall Stark
1
1College of Medical, Veterinary and Life Sciences, University of Glasgow, Bower Building, Glasgow G12 8QQ, U.K.
1To whom correspondence should be addressed (email Marshall.Stark@glasgow.ac.uk).
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Martin R. Boocock;
Martin R. Boocock
1College of Medical, Veterinary and Life Sciences, University of Glasgow, Bower Building, Glasgow G12 8QQ, U.K.
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Femi J. Olorunniji;
Femi J. Olorunniji
1College of Medical, Veterinary and Life Sciences, University of Glasgow, Bower Building, Glasgow G12 8QQ, U.K.
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Sally-J. Rowland
Sally-J. Rowland
1College of Medical, Veterinary and Life Sciences, University of Glasgow, Bower Building, Glasgow G12 8QQ, U.K.
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Publisher: Portland Press Ltd
Received:
September 02 2010
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© The Authors Journal compilation © 2011 Biochemical Society
2011
Biochem Soc Trans (2011) 39 (2): 617–622.
Article history
Received:
September 02 2010
Citation
W. Marshall Stark, Martin R. Boocock, Femi J. Olorunniji, Sally-J. Rowland; Intermediates in serine recombinase-mediated site-specific recombination. Biochem Soc Trans 1 April 2011; 39 (2): 617–622. doi: https://doi.org/10.1042/BST0390617
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