Modification of ionizing radiation clustered damage: estimate of the migration distance of holes through DNA via guanyl radicals under physiological conditions.
OBJECTIVE
Guanyl radicals are produced in DNA when it is subjected to oxidation or ionizing radiation. The sites at which stable products can be identified can be located dozens of base pairs away from the initial site of the electron loss. This migration will modify the spatial distribution of damage and tends to mitigate the clustering of initial damage generally associated with ionizing radiation. The migration distance is presumably a function of the lifetime of the intermediate guanyl radical, and we wished to quantify the relationship between them.


METHODS
Aqueous solutions containing plasmid DNA and thiocyanate ions were treated with gamma-irradiation. These conditions result in the very efficient production of guanyl radicals in the plasmid. We quantified the formation of stable guanine oxidation products in the plasmid as strand breaks by using the E. coli base excision repair endonuclease formamidopyrimidine-DNA N-glycosylase (FPG). The effect of two additives on the yield of guanine oxidation, nitrite ions and the DNA binding ligand doxorubicin (adriamycin), were examined.


RESULTS
The presence during irradiation of the DNA-binding ligand doxorubicin attenuated the yields of stable oxidized guanine products formed. The additional presence of nitrite decreased this effect of doxorubicin.


CONCLUSIONS
Because doxorubicin binds strongly to DNA, its ability to attenuate guanine oxidation can be interpreted in terms of the migration distance of the intermediate guanyl radical. Because nitrite repairs these intermediate guanyl radicals by electron transfer, its presence during irradiation decreases their lifetime. Therefore, we derived an estimate of the migration distance of guanyl radicals as a function of their lifetime. The presence in cells of antioxidants such as glutathione sets an upper limit to the likely lifetime and, therefore, the migration distance of guanyl radicals. It was concluded that the migration of guanyl radicals may not decrease the clustering of DNA damage in vivo to a great extent.
Department of Radiology, University of California at San Diego, La Jolla, CA 92093-0610, USA. jmilligan@ucsd.edu
J R
JR
Milligan
J A
JA
Aguilera
R A
RA
Paglinawan
K J
KJ
Nguyen
J F
JF
Ward
D000970:Antineoplastic Agents / Q000627:therapeutic use; D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000627:therapeutic use; D004926:Escherichia coli / Q000378:metabolism; D005609:Free Radicals; D006147:Guanine / Q000737:chemistry*; D015233:Models, Statistical; D010957:Plasmids / Q000378:metabolism; D013997:Time Factors
D000970:Antineoplastic Agents; D005609:Free Radicals; D006147:Guanine; D004317:Doxorubicin; D004247:DNA