DNA strand breakage by 125I-decay in a synthetic oligodeoxynucleotide--1. Fragment distribution and evaluation of DMSO protection effect.
A double stranded oligodeoxynucleotide containing a single 125I-dC in a defined location was used to investigate DNA strand breakage resulting from 125I decay. Samples of a 41 bp oligodeoxynucleotide were incubated in 20 mM phosphate buffer (PB), or PB plus 2 M dimethylsulphoxide (DMSO), at 4 degrees C during 18-20 days. The 32P-5'-end labelled DNA fragments produced by 125I decays were separated on denaturing polyacrylamide gels, and the 32P activity in each fragment was determined by scintillation counting after elution of fragments from gel. Most of the breaks, around 90%, occurred within 4-5 nucleotides of the 125I-dC, but DNA breaks were detected up to 16 nucleotides from the decay site. The 125I-dC was located at the 21st nucleotide from the 32P-5'-end label, and since 32P was not detected in fragments longer than 20 nucleotides, it was assumed that all 125I decay events produce at least one break in the 125I-labelled DNA strand. The results show a considerable protection effect of DMSO on DNA breaks at sites >5-6 nucleotides from the 125I location. The probability of breaks in this region was decreased with DMSO by a factor of 2 to 8-fold, suggesting significant role for radical-mediated DNA breaks at the more distant sites. However, the total protection effect of DMSO is rather small: 1.1, because of the small contribution of breakage at distant sites to the total yield.
Kandaiya
I S
IS
Department of Molecular Radiation Biology, Peter MacCallum Cancer Institute, Melbourne, Australia.
Lobachevsky
P N
PN
D'Cunha
G
G
Martin
R F
RF
D001483:Base Sequence / Q000528:radiation effects; D004247:DNA / Q000187:drug effects / Q000528:radiation effects*; D004249:DNA Damage; D003854:Deoxyribonucleotides; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D007457:Iodine Radioisotopes; D018499:Linear Energy Transfer; D008969:Molecular Sequence Data
D003854:Deoxyribonucleotides; D007457:Iodine Radioisotopes; D004247:DNA; D004121:Dimethyl Sulfoxide