Formation of single and double strand breaks in DNA ultraviolet irradiated at high intensity.
The induction of single-strand breaks (SSB) by two quantum processes in DNA is well established. We now report that biphotonic processes result in double-strand breaks (DSB) as well. pUC19 and bacteriophage M13 RF DNA were irradiated using an excimer laser (248 nm) at intensities of 10(7), 10(9), 10(10) and 10(11) W/m2 and doses up to 30 kJ/m2. The proportion of DNA as supercoil, open circular, linear and short fragments was determined by gel electrophoresis. Linear molecules were noted at fluences where supercoiled DNA was still present. The random occurrence of independent SSB in proximity to each other on opposite strands (producing linear DNA) implies introduction of numerous SSB per molecule in the sample. If so, supercoiled DNA that has sustained no SSB should not be observed. A model accounting for the amounts of supercoiled, open circular, linear and shorter fragments of DNA due to SSB, DSB and Scissions (opposition of two independently occurring SSB producing an apparent DSB) was developed, our experimental data and those of others were fit to the model, and quantum yields determined for SSB and DSB formation at each intensity. Results showed that high intensity laser radiation caused an increase in the quantum yields for both SSB and DSB formation. The mechanism of DSB formation is unknown, and may be due to simultaneous cleavage of both strands in one biphotonic event or the biased introduction of an SSB opposite a preexisting SSB, requiring two biphotonic events.
Masnyk
T W
TW
Department of Pathology, F. Edward Hébert School of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD 20889-4799.
Minton
K W
KW
D003090:Coliphages / Q000235:genetics; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000528:radiation effects*; D004279:DNA, Viral / Q000528:radiation effects*; D003850:Deoxyribonuclease I; D004307:Dose-Response Relationship, Radiation; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D010957:Plasmids / Q000528:radiation effects*; D014466:Ultraviolet Rays
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D003850:Deoxyribonuclease I