A reliable assessment of 8-oxo-2-deoxyguanosine levels in nuclear and mitochondrial DNA using the sodium iodide method to isolate DNA

Michelle L. Hamilton; Zhong Mao Guo; Clinton D. Fuller; Holly Van Remmen; Walter F. Ward; Steven N. Austad; Dean A. Troyer; Ian Thompson; Arlan Richardson

doi:10.1093/nar/29.10.2117

A reliable assessment of 8-oxo-2-deoxyguanosine levels in nuclear and mitochondrial DNA using the sodium iodide method to isolate DNA

Michelle L. Hamilton, Zhong Mao Guo, Clinton D. Fuller, Holly Van Remmen, Walter F. Ward, Steven N. Austad, Dean A. Troyer, Ian Thompson, Arlan Richardson

Radiation Oncology

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Abstract

A major controversy in the area of DNA biochemistry concerns the actual in vivo levels of oxidative damage in DNA. We show here that 8-oxo-2-deoxyguanosine (oxo8dG) generation during DNA isolation is eliminated using the sodium iodide (Nal) isolation method and that the level of oxo8dG in nuclear DNA (nDNA) is almost one-hundredth of the level obtained using the classical phenol method. We found using Nal that the ratio of oxo8dG/10⁵ deoxyguanosine (dG) in nDNA isolated from mouse tissues ranged from 0.032 ± 0.002 for liver to 0.015 ± 0.003 for brain. We observed a significant increase (10-fold) in oxo8dG in nDNA isolated from liver tissue after 2 Gy of γ-irradiation when Nal was used to isolate DNA. The turnover of oxo8dG in nDNA was rapid, e.g. disappearance of oxo8dG in the mouse liver in vivo after γ-irradiation had a half-life of 11 min. The levels of oxo8dG in mitochondrial DNA isolated from liver, heart and brain were 6-, 16- and 23-fold higher than nDNA from these tissues. Thus, our results showed that the steady-state levels of oxo8dG in mouse tissues range from 180 to 360 lesions in the nuclear genome and from one to two lesions in 100 mitochondrial genomes.

Original language	English (US)
Pages (from-to)	2117-2126
Number of pages	10
Journal	Nucleic acids research
Volume	29
Issue number	10
DOIs	https://doi.org/10.1093/nar/29.10.2117
State	Published - May 15 2001

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Genetics

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title = "A reliable assessment of 8-oxo-2-deoxyguanosine levels in nuclear and mitochondrial DNA using the sodium iodide method to isolate DNA",

abstract = "A major controversy in the area of DNA biochemistry concerns the actual in vivo levels of oxidative damage in DNA. We show here that 8-oxo-2-deoxyguanosine (oxo8dG) generation during DNA isolation is eliminated using the sodium iodide (Nal) isolation method and that the level of oxo8dG in nuclear DNA (nDNA) is almost one-hundredth of the level obtained using the classical phenol method. We found using Nal that the ratio of oxo8dG/105 deoxyguanosine (dG) in nDNA isolated from mouse tissues ranged from 0.032 ± 0.002 for liver to 0.015 ± 0.003 for brain. We observed a significant increase (10-fold) in oxo8dG in nDNA isolated from liver tissue after 2 Gy of γ-irradiation when Nal was used to isolate DNA. The turnover of oxo8dG in nDNA was rapid, e.g. disappearance of oxo8dG in the mouse liver in vivo after γ-irradiation had a half-life of 11 min. The levels of oxo8dG in mitochondrial DNA isolated from liver, heart and brain were 6-, 16- and 23-fold higher than nDNA from these tissues. Thus, our results showed that the steady-state levels of oxo8dG in mouse tissues range from 180 to 360 lesions in the nuclear genome and from one to two lesions in 100 mitochondrial genomes.",

author = "Hamilton, {Michelle L.} and Guo, {Zhong Mao} and Fuller, {Clinton D.} and {Van Remmen}, Holly and Ward, {Walter F.} and Austad, {Steven N.} and Troyer, {Dean A.} and Ian Thompson and Arlan Richardson",

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AU - Hamilton, Michelle L.

AU - Guo, Zhong Mao

AU - Fuller, Clinton D.

AU - Van Remmen, Holly

AU - Ward, Walter F.

AU - Austad, Steven N.

AU - Troyer, Dean A.

AU - Thompson, Ian

AU - Richardson, Arlan

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N2 - A major controversy in the area of DNA biochemistry concerns the actual in vivo levels of oxidative damage in DNA. We show here that 8-oxo-2-deoxyguanosine (oxo8dG) generation during DNA isolation is eliminated using the sodium iodide (Nal) isolation method and that the level of oxo8dG in nuclear DNA (nDNA) is almost one-hundredth of the level obtained using the classical phenol method. We found using Nal that the ratio of oxo8dG/105 deoxyguanosine (dG) in nDNA isolated from mouse tissues ranged from 0.032 ± 0.002 for liver to 0.015 ± 0.003 for brain. We observed a significant increase (10-fold) in oxo8dG in nDNA isolated from liver tissue after 2 Gy of γ-irradiation when Nal was used to isolate DNA. The turnover of oxo8dG in nDNA was rapid, e.g. disappearance of oxo8dG in the mouse liver in vivo after γ-irradiation had a half-life of 11 min. The levels of oxo8dG in mitochondrial DNA isolated from liver, heart and brain were 6-, 16- and 23-fold higher than nDNA from these tissues. Thus, our results showed that the steady-state levels of oxo8dG in mouse tissues range from 180 to 360 lesions in the nuclear genome and from one to two lesions in 100 mitochondrial genomes.

AB - A major controversy in the area of DNA biochemistry concerns the actual in vivo levels of oxidative damage in DNA. We show here that 8-oxo-2-deoxyguanosine (oxo8dG) generation during DNA isolation is eliminated using the sodium iodide (Nal) isolation method and that the level of oxo8dG in nuclear DNA (nDNA) is almost one-hundredth of the level obtained using the classical phenol method. We found using Nal that the ratio of oxo8dG/105 deoxyguanosine (dG) in nDNA isolated from mouse tissues ranged from 0.032 ± 0.002 for liver to 0.015 ± 0.003 for brain. We observed a significant increase (10-fold) in oxo8dG in nDNA isolated from liver tissue after 2 Gy of γ-irradiation when Nal was used to isolate DNA. The turnover of oxo8dG in nDNA was rapid, e.g. disappearance of oxo8dG in the mouse liver in vivo after γ-irradiation had a half-life of 11 min. The levels of oxo8dG in mitochondrial DNA isolated from liver, heart and brain were 6-, 16- and 23-fold higher than nDNA from these tissues. Thus, our results showed that the steady-state levels of oxo8dG in mouse tissues range from 180 to 360 lesions in the nuclear genome and from one to two lesions in 100 mitochondrial genomes.

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A reliable assessment of 8-oxo-2-deoxyguanosine levels in nuclear and mitochondrial DNA using the sodium iodide method to isolate DNA

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