Electron trap for DNA-bound repair enzymes: a strategy for DNA-mediated signaling

Proc Natl Acad Sci U S A. 2006 Mar 7;103(10):3610-4. doi: 10.1073/pnas.0600239103. Epub 2006 Feb 27.

Abstract

Despite a low copy number within the cell, base excision repair (BER) enzymes readily detect DNA base lesions and mismatches. These enzymes also contain [Fe4S4] clusters, yet a redox role for these iron cofactors had been unclear. Here, we provide evidence that BER proteins may use DNA-mediated redox chemistry as part of a signaling mechanism to detect base lesions. By using chemically modified bases, we show electron trapping on DNA in solution with bound BER enzymes by electron paramagnetic resonance (EPR) spectroscopy. We demonstrate electron transfer from two BER proteins, Endonuclease III (EndoIII) and MutY, to modified bases in DNA containing oxidized nitroxyl radical EPR probes. Electron trapping requires that the modified base is coupled to the DNA pi-stack, and trapping efficiency is increased when a noncleavable MutY substrate analogue is located distally to the trap. These results are consistent with DNA binding leading to the activation of the repair proteins toward oxidation. Significantly, these results support a mechanism for DNA repair that involves DNA-mediated charge transport.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Base Sequence
  • DNA Glycosylases / metabolism
  • DNA Repair Enzymes / metabolism*
  • DNA Repair*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism
  • Deoxyribonuclease (Pyrimidine Dimer) / metabolism
  • Electron Spin Resonance Spectroscopy
  • Electron Transport
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / metabolism
  • Models, Biological
  • Signal Transduction
  • Spin Labels

Substances

  • DNA, Bacterial
  • Escherichia coli Proteins
  • Spin Labels
  • Deoxyribonuclease (Pyrimidine Dimer)
  • NTH protein, E coli
  • DNA Glycosylases
  • mutY adenine glycosylase
  • DNA Repair Enzymes