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

Nucleic Acids Res. 1999 Oct 1;27(19):3875-80. doi: 10.1093/nar/27.19.3875.

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 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.

MeSH terms

  • DNA / chemistry
  • DNA / metabolism
  • DNA / ultrastructure*
  • DNA Helicases
  • DNA-Binding Proteins / genetics*
  • Deoxyribodipyrimidine Photo-Lyase / metabolism*
  • Humans
  • Microscopy, Atomic Force
  • Models, Molecular
  • Nucleic Acid Conformation

Substances

  • DNA-Binding Proteins
  • XPBC-ERCC-3 protein
  • DNA
  • DNA Helicases
  • Deoxyribodipyrimidine Photo-Lyase