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| Subject Categories: Genome Stability & Dynamics |
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| The EMBO Journal
(2003) 22, 1398–1409, doi:10.1093/emboj/cdg132 |
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| The complex between a four-way DNA junction and T7 endonuclease I |
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| Anne-Cécile Déclais1, Jonathan M. Fogg1, Alasdair D. J. Freeman1, Franck Coste1, Jonathan M. Hadden2, Simon E. V. Phillips2 and David M. J. Lilley1 |
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1 Cancer Research UK Nucleic Acid Structure Research Group, Department of Biochemistry, MSI/WTB Complex, The University of Dundee, Dundee DD1 5EH, UK 2 Astbury Centre for Structural Molecular Biology, School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK To whom correspondence should be addressed David M. J. Lilley, d.m.j.lilley@dundee.ac.uk Received 23 October 2002; Revised 20 January 2003; Accepted 27 January 2003. |
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| Abstract |
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| The junction-resolving enzyme endonuclease I is selective for the structure of the DNA four-way (Holliday) junction. The enzyme binds to a four-way junction in two possible orientations, with a 4:1 ratio, opening the DNA structure at the centre and changing the global structure into a 90° cross of approximately coaxial helices. The nuclease cleaves the continuous strands of the junction in each orientation. Binding leads to pronounced regions of protection of the DNA against hydroxyl radical attack. Using all this information together with the known structure of the enzyme and the structure of the BglI–DNA complex, we have constructed a model of the complex of endonuclease I and a DNA junction. This shows how the enzyme is selective for the structure of a four-way junction, such that both continuous strands can be accommodated into the two active sites so that a productive resolution event is possible. |
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| Keywords: DNA repair, Holliday junction, nucleases, recombination |
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