Modified alkaline elution allows the measurement of intact apurinic sites in mammalian genomic DNA.
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.
Malvy
C
C
UMR 8532 CNRS, Physicochimie et Pharmacologie des Macromolécules Biologiques, Institut Gustave Roussy, rue Camille-Desmoulins, 94800, Villejuif, France. cmalvy@igr.fr
Lefrançois
M
M
Bertrand
J R
JR
Markovits
J
J
D000818:Animals; D001080:Apurinic Acid / Q000737:chemistry* / Q000302:isolation & purification; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007939:Leukemia L1210 / Q000378:metabolism / Q000473:pathology; D008322:Mammals; D051379:Mice; D009842:Oligopeptides; D051381:Rats; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D009842:Oligopeptides; D001080:Apurinic Acid; C029535:lysyl-tryptophyl-alpha-lysine; D004247:DNA