Single-Strand Breaks in Oligodeoxyribonucleotides Induced by Fission Neutrons and Gamma Radiation and Measured by Gel Electrophoresis

Protective Effects of Aminothiols

Charles E. Swenberg, Yashesh N. Vaishnav, Bin Li, Hong Tsao, Bing Mao, Nicholas Geacintov

Research output: Contribution to journalArticle

Abstract

The technique of high-resolution gel electrophoresis using oligodeoxyribonucleotides of known composition as model systems, offers a simple quantitative estimate of DNA damage in aqueous solution induced by ionizing radiation. The fraction of damaged DNA can be quantitatively defined in terms of the increased electrophoretic mobilities of the damaged oligonucleotides, relative to the mobility of the unirradiated and intact oligonucleotides. The usual direct strand breaks can be observed at γ-ray dosages of 200 Gy. However, at a γ-ray dosage of 400 Gy, only a broad background, attributed to heterogeneously and multiply damaged oligonucleotide fragments with overlapping and varying electrophoretic mobilities, can be distinguished. On the other hand, individual bands due to resolvable DNA fragments are evident even at dosages as high as 400 Gy for fission neutrons. When double-stranded oligonucleotides are exposed to γ-ray dosages of 200 Gy, the fraction of damaged DNA approaches 30-40%. This damage can be almost completely suppressed (>99%) if the irradiations are conducted in aqueous solutions in the presence of 0.5-1.0 mM concentrations of the thiols cysteamine or 3-(3-methylaminopropylamino)propanethiol (WR-151326). The rate constant of reaction of OH· radicals with small double stranded oligonucleotides 16 base pairs long, KDNA, is found to be closer to the diffusion-controlled value (> 3 × 109 M-1 s-1) than the magnitudes of KDNA for the higher molecular weight, native DNA reported in the literature. These observations suggest that oligonucleotides represent more simple model systems than native DNA in solutions for studying the mechanisms of radioprotection exerted by thiols of different structures.

Original languageEnglish (US)
Pages (from-to)241-254
Number of pages14
JournalJournal of Radiation Research
Volume38
Issue number4
StatePublished - Dec 1997

Fingerprint

oligodeoxyribonucleotides
oligonucleotides
Oligodeoxyribonucleotides
Gamma Rays
Neutrons
electrophoresis
Oligonucleotides
strands
gel electrophoresis
gamma radiation
fission
protective effect
Electrophoresis
deoxyribonucleic acid
Gels
gels
gamma rays
neutrons
radiation
DNA

Keywords

  • Cysteamine
  • DNA oligomers
  • Gel electrophoresis
  • Radiation protection
  • Strand breaks

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Radiology Nuclear Medicine and imaging
  • Radiation
  • Radiological and Ultrasound Technology

Cite this

Single-Strand Breaks in Oligodeoxyribonucleotides Induced by Fission Neutrons and Gamma Radiation and Measured by Gel Electrophoresis : Protective Effects of Aminothiols. / Swenberg, Charles E.; Vaishnav, Yashesh N.; Li, Bin; Tsao, Hong; Mao, Bing; Geacintov, Nicholas.

In: Journal of Radiation Research, Vol. 38, No. 4, 12.1997, p. 241-254.

Research output: Contribution to journalArticle

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abstract = "The technique of high-resolution gel electrophoresis using oligodeoxyribonucleotides of known composition as model systems, offers a simple quantitative estimate of DNA damage in aqueous solution induced by ionizing radiation. The fraction of damaged DNA can be quantitatively defined in terms of the increased electrophoretic mobilities of the damaged oligonucleotides, relative to the mobility of the unirradiated and intact oligonucleotides. The usual direct strand breaks can be observed at γ-ray dosages of 200 Gy. However, at a γ-ray dosage of 400 Gy, only a broad background, attributed to heterogeneously and multiply damaged oligonucleotide fragments with overlapping and varying electrophoretic mobilities, can be distinguished. On the other hand, individual bands due to resolvable DNA fragments are evident even at dosages as high as 400 Gy for fission neutrons. When double-stranded oligonucleotides are exposed to γ-ray dosages of 200 Gy, the fraction of damaged DNA approaches 30-40{\%}. This damage can be almost completely suppressed (>99{\%}) if the irradiations are conducted in aqueous solutions in the presence of 0.5-1.0 mM concentrations of the thiols cysteamine or 3-(3-methylaminopropylamino)propanethiol (WR-151326). The rate constant of reaction of OH· radicals with small double stranded oligonucleotides 16 base pairs long, KDNA, is found to be closer to the diffusion-controlled value (> 3 × 109 M-1 s-1) than the magnitudes of KDNA for the higher molecular weight, native DNA reported in the literature. These observations suggest that oligonucleotides represent more simple model systems than native DNA in solutions for studying the mechanisms of radioprotection exerted by thiols of different structures.",
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