Modified alkaline elution allows the measurement of intact apurinic sites in mammalian genomic DNA

Biochimie. 2000 Aug;82(8):717-21. doi: 10.1016/s0300-9084(00)01152-4.

Abstract

The presence of apurinic/apyrimidinic (AP) sites in cell genomes is known to be toxic and mutagenic. These lesions are therefore repaired in cells by efficient enzymatic systems. However, a report (Nakamura and Swenberg, Cancer Res. 59 (1999) 2522-2526) indicates an unexpected high rate of endogenous apurinic/apyrimidinic (AP) sites in genomic DNA in mammalian tissues. The technology used does not allow the authors to distinguish between intact AP sites and 3'cleaved AP sites. The corresponding values range between 2 and 4 sites per million of nucleotides in various human and rat tissues. Using a modified alkaline elution method we show here that the stationary level of intact AP sites is about 0.16 per million of nucleotides in leukemic mouse L1210 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apurinic Acid / chemistry*
  • Apurinic Acid / isolation & purification
  • DNA / chemistry*
  • DNA / isolation & purification
  • DNA Damage*
  • DNA, Neoplasm / chemistry*
  • DNA, Neoplasm / isolation & purification
  • Humans
  • Hydrogen-Ion Concentration
  • Kinetics
  • Leukemia L1210 / metabolism
  • Leukemia L1210 / pathology
  • Mammals
  • Mice
  • Oligopeptides
  • Rats
  • Tumor Cells, Cultured

Substances

  • DNA, Neoplasm
  • Oligopeptides
  • Apurinic Acid
  • lysyl-tryptophyl-alpha-lysine
  • DNA