The Physical Measurement of Radiation Damage to Coliphage T7 DNA

Abstract

Coliphage T7 was exposed to (60)Co gamma radiation while suspended in phosphate buffer or in phosphate buffer plus 0.001 M l-histidine. DNA was isolated from the phage by incubation with pronase, followed by extraction with cold phenol. The intrinsic viscosity of the DNA was measured as a function of radiation dose. The fraction of DNA molecules surviving radiation treatment with no double-strand breaks was measured from the radiation-induced heterogeneity of the DNA sedimentation boundary. From comparison of these measurements it is concluded that radiation introduces lesions other than double-strand breaks which affect the hydrodynamic properties of the DNA. In both buffer and buffer plus histidine the surviving fraction of intact virus genomes far exceeds the surviving fraction of plaque-forming units at any given dose. It was found that the decrease in intrinsic viscosity with dose is independent of the presence of histidine in the radiation medium. From this it is concluded that DNA damage is primarily due to a direct effect of radiation on the phage particle. The procedure necessary to isolate DNA from irradiated virus suggests that radiation produces covalent bonding of protein to the DNA.