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Nucleic Acids Research, Vol 27, Issue 19 3875-3880, Copyright © 1999 by Oxford University Press


ARTICLES

DNA bending by photolyase in specific and non-specific complexes studied by atomic force microscopy

J van Noort, F Orsini, A Eker, C Wyman, B de Grooth and J Greve
Applied Optics Group, Department of Applied Physics, University of Twente, PO Box 217, 7500 AE Enschede, The Netherlands.

Specific and non-specific complexes of DNA and photolyase are visualised by atomic force microscopy. As a substrate for photolyase a 1150 bp DNA restriction fragment was UV-irradiated to produce damaged sites at random positions. Comparison with a 735 bp undamaged DNA fragment made it possible to separate populations of specific and non- specific photolyase complexes on the 1150 bp fragment, relieving the need for highly defined substrates. Thus it was possible to compare DNA bending for specific and non-specific interactions. Non-specific complexes show no significant bending but increased rigidity compared to naked DNA, whereas specific complexes show DNA bending of on average 36 degrees and higher flexibility. A model obtained by docking shows that photolyase can accommodate a 36 degrees bent DNA in the vicinity of the active site.
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