Analysis of DNA strand cleavage at abasic sites

Walter A. Deutsch, Vijay Hegde

Research output: Contribution to journalArticle

Abstract

Abasic sites in DNA arise under a variety of circumstances, including destabilization of bases through oxidative stress, as an intermediate in base excision repair, and through spontaneous loss. Their persistence can yield a blockade to RNA transcription and DNA synthesis and can be a source of mutations. Organisms have developed an enzymatic means of repairing abasic sites in DNA that generally involves a DNA repair pathway that is initiated by a repair protein creating a phosphodiester break (“nick”) adjacent to the site of base loss. Here we describe a method for analyzing the manner in which repair endonucleases differ in the way they create nicks in DNA and how to distinguish between them using cellular crude extracts.

Original languageEnglish
Pages (from-to)159-168
Number of pages10
JournalMethods in Molecular Biology
Volume1105
DOIs
StatePublished - 2014

Keywords

  • AP endonucleases
  • AP lyases
  • Base excision repair
  • DNA abasic sites
  • DNA damage
  • DNA oligonucleotides
  • Oxidative stress
  • Tetrahydrofuran

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