Cu(II)/H2O2-induced DNA damage is enhanced by packaging of DNA as a nucleosome.

摘要

Copper is a physiologically important, redox-active metal that may be involved in endogenous DNA damage and mutagenesis. To understand the factors that affect the location and quantity of copper-induced oxidative DNA damage in cells, we used the 5S rDNA nucleosome as a model to assess the effect of chromatin structure on DNA damage produced by Cu(II)/H2O2. Packaging of DNA into a nucleosome increased the extent of Cu(II)/H2O2-induced strand breaks by a factor of 2, while the extent of base lesions sensitive to Fpg and endo III glycosylases increased 8-fold. We also observed that Cu(II)/H2O2 caused slightly more strand breaks than base lesions in isolated 5S rDNA (ratio of base lesions to strand breaks of approximately 0.6), while base lesions outnumbered strand breaks by a factor of 3-4 when the DNA was incorporated into a nucleosome. Apart from several sites of enhanced or diminished DNA damage, there were no major changes in the sequence selectivity of Cu(II)/H2O2, and there was no apparent footprinting effect associated with nucleosome structure, such as that observed with the Fe(II)-EDTA complex. Possible mechanisms for explaining these observations include (1) an increase in Cu(II) concentration in the vicinity of nucleosomal DNA caused by binding of Cu to histone proteins or (2) increased reactivity or accessibility of nucleobases caused by DNA conformational changes associated with nucleosome structure. The enhancement of Cu(II)/H2O2-induced DNA damage in nucleosomes stands in contrast to the protective effect afforded DNA by proteins in chromatin against radiation-induced DNA damage.