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Nucleic Acids Research Advance Access originally published online on October 5, 2006
Nucleic Acids Research 2006 34(19):5577-5584; doi:10.1093/nar/gkl447
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Nucleic Acids Research, 2006, Vol. 34, No. 19 5577-5584
© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Structural Biology

RusA Holliday junction resolvase: DNA complex structure—insights into selectivity and specificity

Rachel Macmaster, Svetlana Sedelnikova, Patrick J. Baker, Edward L. Bolt1, Robert G. Lloyd1 and John B. Rafferty*

Department of Molecular Biology and Biotechnology, University of Sheffield Firth Court, Western Bank, Sheffield S10 2TN, UK 1 Institute of Genetics, University of Nottingham Queen's Medical Centre, Nottingham NG7 2UH, UK

*To whom correspondence should be addressed. Tel: +44 114 222 2809; Fax: +44 114 222 2800; Email: j.rafferty{at}sheffield.ac.uk

Received March 30, 2006. Revised June 8, 2006. Accepted June 9, 2006.

We have determined the structure of a catalytically inactive D70N variant of the Escherichia coli RusA resolvase bound to a duplex DNA substrate that reveals critical protein–DNA interactions and permits a much clearer understanding of the interaction of the enzyme with a Holliday junction (HJ). The RusA enzyme cleaves HJs, the fourway DNA branchpoints formed by homologous recombination, by introducing symmetrical cuts in the phosphodiester backbone in a Mg2+ dependent reaction. Although, RusA shows a high level of selectivity for DNA junctions, preferring to bind fourway junctions over other substrates in vitro, it has also been shown to have appreciable affinity for duplex DNA. However, RusA does not show DNA cleavage activity with duplex substrates. Our structure suggests the possible basis for structural selectivity as well as sources of the sequence specificity observed for DNA cleavage by RusA.


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