New methods for detection of low levels of DNA damage in human populations.
The use of a postlabeling method to characterize and to detect infrequent base modifications in DNA is outlined. This method has the advantage that low levels of DNA modifications, approximately 1 modified base per 10(5) nucleotides, can be detected. Moreover, a broad spectrum of modification can be identified by using this methodology. The basis for the method involves transfer of a radioactive phosphate from the gamma position of ATP to the 5'-hydroxyl terminus of 3'-phosphoryl nucleotides that are derived from modified DNA by appropriate nuclease digestion. The second method involves use of a defined DNA sequence within human cells. The alpha sequence is used as a probe for DNA damage to specific nucleotides. The alpha DNA sequence is reiterated approximately 300,000 times in the human genome and exists in tandem arrays. It comprises approximately 1% of the entire genome. The reiterated sequence is sufficiently homogeneous to permit its use as a probe for a site specific in DNA damage. Examples of the application of both of these methodologies to DNA damage inflicted in human cells by chemicals and ultraviolet light are provided.
D000477:Alkylating Agents / Q000494:pharmacology; D001483:Base Sequence / Q000187:drug effects / Q000528:radiation effects; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis*; D006801:Humans; D009152:Mutagenicity Tests / Q000379:methods*; D010761:Phosphorus Radioisotopes; D013463:Sulfuric Acid Esters / Q000494:pharmacology; D014466:Ultraviolet Rays
D000477:Alkylating Agents; D010761:Phosphorus Radioisotopes; D013463:Sulfuric Acid Esters; D004247:DNA; C007482:dimethyl sulfate