Misincorporation of uracil into the DNA of folate- and B12-deficient HL60 cells.
HL60 cells were cultured for 10 days under various experimental conditions. They were then incubated with 1 mumol/l [5-3H] uridine for 2 hours and their DNA extracted. The DNA was hydrolysed to deoxyribonucleosides with phosphodiesterase and alkaline phosphatase and the hydrolysate subjected to Aminex A6 chromatography. The elution profiles showed that, when compared with control cells, DNA from cells grown in medium deficient in folate, B12 or both folate and B12 contained increased amounts of deoxyuridine (dU) and increased radioactivity in the dU peak. The data demonstrate that misincorporation of uracil into DNA occurs in a myeloid cell line cultured in growth medium deficient in folate, B12 or both folate and B12.
Wickramasinghe
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Dept. of Haematology, St. Mary's Hospital Medical School, Imperial College of Science, Technology & Medicine, London, U.K.
Fida
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D002852:Chromatography, Ion Exchange; D004273:DNA, Neoplasm / Q000096:biosynthesis* / Q000187:drug effects / Q000302:isolation & purification; D005492:Folic Acid / Q000494:pharmacology*; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute; D008727:Methotrexate / Q000494:pharmacology; D014316:Tritium; D014407:Tumor Cells, Cultured; D014498:Uracil / Q000378:metabolism*; D014805:Vitamin B 12 / Q000494:pharmacology*
D004273:DNA, Neoplasm; D014316:Tritium; D014498:Uracil; D005492:Folic Acid; D014805:Vitamin B 12; D008727:Methotrexate