Searching for DNA lesions: structural evidence for lower- and higher-affinity DNA binding conformations of human alkyladenine DNA glycosylase

Biochemistry. 2012 Jan 10;51(1):382-90. doi: 10.1021/bi201484k. Epub 2011 Dec 20.

Abstract

To efficiently repair DNA, human alkyladenine DNA glycosylase (AAG) must search the million-fold excess of unmodified DNA bases to find a handful of DNA lesions. Such a search can be facilitated by the ability of glycosylases, like AAG, to interact with DNA using two affinities: a lower-affinity interaction in a searching process and a higher-affinity interaction for catalytic repair. Here, we present crystal structures of AAG trapped in two DNA-bound states. The lower-affinity depiction allows us to investigate, for the first time, the conformation of this protein in the absence of a tightly bound DNA adduct. We find that active site residues of AAG involved in binding lesion bases are in a disordered state. Furthermore, two loops that contribute significantly to the positive electrostatic surface of AAG are disordered. Additionally, a higher-affinity state of AAG captured here provides a fortuitous snapshot of how this enzyme interacts with a DNA adduct that resembles a one-base loop.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Catalysis
  • Crystallography, X-Ray
  • DNA Adducts / chemistry
  • DNA Damage*
  • DNA Glycosylases / chemistry*
  • DNA Glycosylases / genetics*
  • DNA Glycosylases / metabolism
  • DNA Repair
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Genome, Human
  • Humans
  • Mutagenesis
  • Nucleic Acid Heteroduplexes / chemistry
  • Nucleic Acid Heteroduplexes / genetics
  • Nucleic Acid Heteroduplexes / metabolism
  • Plasmids
  • Protein Conformation

Substances

  • DNA Adducts
  • DNA-Binding Proteins
  • Nucleic Acid Heteroduplexes
  • 3-methyladenine-DNA glycosylase
  • DNA Glycosylases

Associated data

  • PDB/1BNK
  • PDB/1F4R
  • PDB/3QI5
  • PDB/3UBY