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° and higher flexibility. A model obtained by docking shows that photolyase can accommodate a 36° bent DNA in the vicinity of the active site.
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, J. Greve. - In: NUCLEIC ACIDS RESEARCH. - ISSN 0305-1048. - 27:19(1999), pp. 3875-3880. [10.1093/nar/27.19.3875]
DNA bending by photolyase in specific and non-specific complexes studied by atomic force microscopy
F. Orsini;
1999
Abstract
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° and higher flexibility. A model obtained by docking shows that photolyase can accommodate a 36° bent DNA in the vicinity of the active site.Pubblicazioni consigliate
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