Misincorporation of Deoxyuridine in Human Cells: Consequences of Antifolate Exposure

  • Randall G. Richards
  • Oliver E. Brown
  • W. David Sedwick
Chapter
Part of the Basic Life Sciences book series (BLSC, volume 31)

Abstract

Treatment of B and T lymphoblastoid cell lines (SB and MOLT-4, respectively) and a promyelocytic leukemia cell line (HL-60) with the lipid soluble antifolate, 2,4-diamino-5-methyl-6-(2’,5’-dimethoxybenzyl)-pyrido(2,4-d) pyrimidine (BW301U), led to drug dose-dependent inhibition of [3H]deoxyuridine (dU) incorporation into DNA as thymidine, and to misincorporation of [3H]dU as dUMP. After a 15 min preincubation with up to 50 μM BW301U and a further 15 min incubation after addition of [3H]dU, the number of alkaline labile apyrimidinic sites increased with increasing drug dose, as demonstrated by alkaline sucrose gradient analysis. Significantly, new replication of DNA was inhibited only ∿50% by 50 μM BW301U when [3H]dU incorporation was ≥97% inhibited. Additional preliminary findings suggest that newly replicated DNA containing misincorporated dUMP is rapidly degradedin vivo by extensive excision-repair processes.

Keywords

Replication Fork Lymphoblastoid Cell Line Micrococcal Nuclease Nucleus Isolation Promyelocytic Leukemia Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Randall G. Richards
    • 1
  • Oliver E. Brown
    • 1
  • W. David Sedwick
    • 1
  1. 1.Departments of Medicine and Microbiology-ImmunologyDuke University Medical CenterDurhamUSA

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