Analysis of DNA strand cleavage at abasic sites.
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.
Deutsch
Walter A
WA
Pennington Biomedical Research Center, Louisiana State University, Baton Rouge, LA, USA.
Hegde
Vijay
V
D000818:Animals; D001080:Apurinic Acid / Q000032:analysis; D004247:DNA / Q000737:chemistry; D004249:DNA Damage; D004260:DNA Repair; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000737:chemistry / Q000378:metabolism*; D043224:Deoxyribonuclease (Pyrimidine Dimer) / Q000737:chemistry; D004330:Drosophila / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry; D005663:Furans / Q000737:chemistry; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011119:Polynucleotides / Q000032:analysis
D029968:Escherichia coli Proteins; D005663:Furans; D009838:Oligodeoxyribonucleotides; D011119:Polynucleotides; C001790:apyrimidinic acid; D001080:Apurinic Acid; C018674:tetrahydrofuran; D004247:DNA; D043224:Deoxyribonuclease (Pyrimidine Dimer); C106403:NTH protein, E coli; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase