Diaminopurine-resistant mutants of cultured, diploid human fibroblasts.
Clones of cells resistant to 2,6-diaminopurine were detected in skin fibroblast cultures derived from 13 of 21 normal humans of both sexes from 17 unrelated families. Almost all of the cultures that yielded mutants were chosen for further study from among a total of 83 surveyed because they displayed a slight resistance to low concentrations of diaminopurine. The incidences of mutant colonies ranged between about 10(-5) and 10(-4) per cell surviving prior mutagenic treatment with MNNG. The incidences of spontaneous mutants were about 10(-7) to 10(-5) in three unrelated cultures. Most independent mutants had distinctly reduced activity of adenine phosphoribosyltransferase but some had apparently normal amounts of activity. Two mutants from unrelated boys had little or no detectable enzyme activity and were unable to effectively use exogenous adenine for growth when purine biosynthesis was blocked with azaserine. Most mutants could utilize exogenous adenine, just as most azaguanine-resistant fibroblast mutants can utilize exogenous hypoxanthine, even when their hypoxanthine-guanine phosphoribosyltransferase activity is reduced. Diverse genetic changes conferred diaminopurine resistance but their specific natures are still undefined. Gross numerical or structural chromosome abnormalities were not observed in the mutants examined so far. Since at least one gene responsible for adenine phosphoribosyltransferase activity is on autosome No. 16 our results suggest that at least some of the cultures yielding mutants were heterozygous and that alleles conferring diaminopurine resistance may be frequent enough to comprise a polymorphism.
D000225:Adenine; D002250:Carbon Radioisotopes; D002999:Clone Cells; D003470:Culture Media; D003959:Diamines / Q000494:pharmacology; D004171:Diploidy; D004351:Drug Resistance; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006579:Heterozygote; D006801:Humans; D007292:Inosine Nucleotides; D008297:Male; D009154:Mutation; D010375:Pedigree; D010428:Pentosephosphates; D010430:Pentosyltransferases / Q000378:metabolism; D010756:Phosphoric Acids; D011110:Polymorphism, Genetic; D011687:Purines / Q000494:pharmacology*; D012867:Skin / Q000187:drug effects / Q000378:metabolism*
D002250:Carbon Radioisotopes; D003470:Culture Media; D003959:Diamines; D007292:Inosine Nucleotides; D010428:Pentosephosphates; D010756:Phosphoric Acids; D011687:Purines; D010430:Pentosyltransferases; D000225:Adenine