Journal Article

A study of endonuclease III-sensitive sites in irradiated DNA: detection of α-particle-induced oxidative damage

Kevin M. Prise, Clare H.L. Pullar and Barry D. Michael

in Carcinogenesis

Volume 20, issue 5, pages 905-909
Published in print May 1999 | ISSN: 0143-3334
Published online May 1999 | e-ISSN: 1460-2180 | DOI: http://dx.doi.org/10.1093/carcin/20.5.905
A study of endonuclease III-sensitive sites in irradiated DNA: detection of α-particle-induced oxidative damage

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An important difference between chemical agents that induce oxidative damage in DNA and ionizing radiation is that radiation-induced damage is clustered locally on the DNA. Both modelling and experimental studies have predicted the importance of clustering of lesions induced by ionizing radiation and its dependence on radiation quality. With increasing linear energy transfer, it is predicted that complex lesions will be formed within 1–20 bp regions of the DNA. As well as strand breaks, these sites may contain multiple damaged bases. We have compared the yields of single strand breaks (ssb) and double strand breaks (dsb) along with those produced by treatment of irradiated DNA with the enzyme endonuclease III, which recognizes a number of oxidized pyrimidines in DNA and converts them to strand breaks. Plasmid DNA was irradiated under two different scavenging conditions to test the involvement of OH˙ radicals with either 60Co γ-rays or α-particles from a 238Pu source. Under low scavenging conditions (10 mM Tris) γ-irradiation induced 7.1×10–7 ssb Gy/bp, which increased 3.7-fold to 2.6×10–6 ssb Gy/bp with endo III treatment. In contrast the yields of dsb increased by 4.2-fold from 1.5×10–8 to 6.3×10–8 dsb Gy/bp. This equates to an additional 2.5% of the endo III-sensitive sites being converted to dsb on enzyme treatment. For α-particles this increased to 9%. Given that endo III sensitive sites may only constitute ~40% of the base lesions induced in DNA, this suggests that up to 6% of the ssb measured in X- and 22% in α-particle-irradiated DNA could have damaged bases associated with them contributing to lesion complexity.

Keywords: BSA, bovine serum albumin; dsb, double strand breaks; endo III, endonuclease III; FPG, formamidopyrimidine–DNA N-glycosylase; GC–MS, gas chromatography–mass spectrometry; LET, linear energy transfer; RBE, relative biological effectiveness; ssb, single strand breaks.

Journal Article.  4098 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

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