PMID: 293658Nov 1, 1979Paper

Endonucleolytic activity directed towards 8-(2-hydroxy-2-propyl) purines in double-stranded DNA

Proceedings of the National Academy of Sciences of the United States of America
Z LivnehJ Sperling

Abstract

Photoalkylation of circular covalently closed DNA from phage PM2 with isopropyl alcohol by using a free radical photoinitiator and UV light of lambda greater than 305 nm led to the specific 8-substitution of purine moieties in the DNA, yielding 8-(2-hydroxy-2-propyl)adenine and 8-(2-hydroxy-2-propyl)guanine as the only detectable damage in the DNA. Using this specifically photoalkylated DNA as a substrate, we discovered in extracts of Micrococcus luteus an endonucleolytic activity that is directed towards 8-(2-hydroxy-2-propyl) purines in DNA. The activity is not a combination of a DNA-glycosylase and an apurinic site endonuclease. It is not inhibited by single-stranded DNA, by UV- or gamma-irradiated single-stranded DNA, or by normal or depurinated double-stranded DNA. however, gamma- or UV-(254 nm) irradiated double-stranded DNAs to inhibit the activity, hinting at the possibility of a common type of lesion in these damaged DNAs. Divalent cations are not required for the incising activity, and it is fully active in 1 mM EDTA, whereas caffeine and ATP cause inhibition. Extracts of mutant M. luteus lacking pyrimidine-dimer-directed endonucleases were found to contain the endonucleolytic activity in levels comparable to those pr...Continue Reading

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Citations

Aug 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·J C SutherlandP Z Takacs
Aug 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·N J DukerD E Fishbein
Apr 1, 1986·Experimental and Molecular Pathology·N J Duker, P E Gallagher
Feb 1, 1985·International Journal of Radiation Biology and Related Studies in Physics, Chemistry, and Medicine·J Cadet, M Berger

Related Concepts

Alkylation
Alkaline DNase
Base Excision Repair
DNA, Circular
Endonuclease
Micrococcus
Purines
Structure-Activity Relationship
Substrate Specificity

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