Published online 1 June 2004
Nucleic Acids Research, 2004, Vol. 32, No. 10 3017-3027
© 2004 Oxford University Press
Definitions and analysis of DNA Holliday junction geometry
Department of Biochemistry and Biophysics, ALS 2011, Oregon State University, Corvallis, OR 97331-7305, USA
*To whom correspondence should be addressed. Tel: +1 541 737 2769; Fax: +1 541 737 0481; Email: hops{at}onid.orst.edu
Received March 18, 2004; Revised and Accepted May 11, 2004
A number of single-crystal structures have now been solved of the four-stranded antiparallel stacked-X form of the Holliday junction. These structures demonstrate how base sequence, substituents, and drug and ion interactions affect the general conformation of this recombination intermediate. The geometry of junctions had previously been described in terms of a specific set of parameters that include: (i) the angle relating the ends of DNA duplexes arms of the junction (interduplex angle); (ii) the relative rotation of the duplexes about the helix axes of the stacked duplex arms (Jroll); and (iii) the translation of the duplexes along these helix axes (Jslide). Here, we present a consistent set of definitions and methods to accurately calculate each of these parameters based on the helical features of the stacked duplex arms in the single-crystal structures of the stacked-X junction, and demonstrate how each of these parameters contributes to an overall conformational feature of the structure. We show that the values for these parameters derived from global rather than local helical axes through the stacked bases of the duplex arms are the most representative of the stacked-X junction conformation. In addition, a very specific parameter (Jtwist) is introduced which relates the relative orientation of the stacked duplex arms across the junction which, unlike the interduplex angle, is length independent. The results from this study provide a general means to relate the geometric features seen in the crystal structures to those determined in solution.
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