Cell Biology and Toxicology

, Volume 1, Issue 2, pp 75–86 | Cite as

Elevation of dCTP pools in xeroderma pigmentosum variant human fibroblasts alters the effects of DNA repair arrest by arabinofuranosyl cytosine

  • William C. Dunn
  • James D. Regan
  • Ronald D. Snyder
Article
  • 14 Downloads

Abstract

DNA excision repair inhibition by arabinofuranosyl cytosine (ara-C) or by ara-C/hydroxyurea (HU) was measured in log phase and confluent cultures of normal and xeroderma pigmentosium (XP)-variant human fibroblasts following insult by ultraviolet (UV) light (20 J/m2). Repair inhibition was determined by measuring the accumulation of DNA single-strand breaks/108 daltons following cell culture exposure to ara-C or ara-C/HU in a series of 3 hr. pulses up ro 24 hr. after UV insult. Both normal and XP-variant derived cells showed a wide range of sensitivity to ara-C in log phase cells (0.2–9.4 breaks/108 daltons DNA), although strand break accumulation was constant for each specific cell line. The same cells were more sensitive to ara-C/HU with a 2–14 fold increase in DNA strand breaks depending upon the cell line assayed. In confluent cultures of normal cells, maximum sensitivity to ara-C and ara-C/HU was achieved with similar levels of repair inhibition observed (16.1 and 16.5 breaks/108 daltons, respectively). The same level of repair inhibition was observed in confulent XP-variants receiving ara-C/HU, but was reduced by 62–68% in cells treated with ara-C alone. Ara-C repair arrest was more rapidly reversed by competing concentrations of exogenous deoxycytidine (dCyd) in XP-variant compared to normal cells, especially in confluent cell cultures. In ara-C/HU treated cells, the level of dCyd reversal was reduced in the XP-variant when compared to cells exposed to ara-C alone. However, the same addition of HU had relatively little effect on dCyd reversal in normal cells. The measurements of dNTP levels indicate an elevated level of intracellular deoxycytosine triphosphate in XP-variant vs normal cells. The implications of these results are discussed as they relate to possible excision repair anomalies in the XP-variant.

Key words

ara-C dCTP pools DNA repair arrest XP-variant 

Abbreviations

ara-C

arabinofuranosul cytosine

dCTP

deoxycytosine triphosphate

dCyd

deoxycytidine

dNTP

deoxynucleoside triphosphate

dT

thymidine

HU

hydroxyurea

XP

xeroderma pigmentosium

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Copyright information

© Princeton Scientific Publishers, Inc 1985

Authors and Affiliations

  • William C. Dunn
    • 1
  • James D. Regan
    • 1
  • Ronald D. Snyder
    • 2
  1. 1.Biology DivisionOak Ridge National LaboratoryTennessee
  2. 2.Stauffer Chemical CompanyFarmington, Connecticut

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