S1 Nuclease-sensitive Sites in Yeast DNA: An Assay for Radiation-induced Base Damage

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Abstract
DNA has been isolated from γ-irradiated yeast cells and then treated with the enzyme S1 nuclease. This enzyme cleaves DNA specifically at sites where localized denaturation has occurred and can therefore be used as a lesion probe to identify regions in the DNA where base-pairing has been disrupted. By analysing the number of single strand breaks, double strand breaks and alkali-labile sites in the DNA before and after treatment with S1 nuclease, it has been possible to calculate the number of S1 nuclease-sensitive sites induced as a result of exposure to ionizing radiation. These sites were found to occur with a frequency about twice that of the double strand break and are thought to result from a primary ionization event in the DNA.

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