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Effects of Glutathione on Chromium-induced DNA Crosslinking and DNA Polymerase Arrest

  • Chapter
Molecular Mechanisms of Metal Toxicity and Carcinogenesis

Part of the book series: Developments in Molecular and Cellular Biochemistry ((DMCB,volume 34))

Abstract

Hexavalent chromium (Cr (VI)) is reduced intracellularly to Cr (V), Cr (IV) and Cr (HI) by ascorbate (Asc), cysteine and glutathione (GSH). These metabolites induce a spectrum of genomic DNA damage resulting in the inhibition of DNA replication. Our previous studies have shown that treatment of DNA with Cr (III) or Cr (VI) plus Asc results in the formation of DNA-CrDNA crosslinks (Cr-DDC) and guanine-specific arrests of both prokaryotic and mammalian DNA polymerases. GSH not only acts as a reductant of Cr (VI) but also becomes crosslinked to DNA by Cr, thus, the focus of the present study was to examine the role of GSH in Cr-induced DNA damage and polymerase arrests. Co-incubation of Cr (III) with plasmid DNA in the presence of GSH led to the crosslinking of GSH to DNA. GSH co-treatment with Cr (III) also led to a decrease in the degree of Cr-induced DNA interstrand crosslinks relative to Cr(III)alone, without affecting total Cr DNA binding. DNA polymerase arrests were observed following treatment of DNA with Cr (III) alone, but were markedly reduced when GSH was added to the reaction mixture. Pre-formed polymerase-arresting lesions (Cr-DDC) were not removed by subsequent addition of GSH. Treatment of DNA with Cr (VI), in the presence of GSH, resulted in crosslinking of GSH to DNA, but failed to produce detectable DNA interstrand crosslinks or polymerase arrests. The inhibitory effect of GSH on Cr-induced polymerase arrest was further confirmed in human genomic DNA using quantitative PCR (QPCR) analysis. Treatment of genomic DNA with Cr (III) resulted in a marked inhibition of the amplification of a 1.6 kb target fragment of the p53 gene byTagpolymerase. This was almost completely prevented by co-treatment with GSH and Cr (III). These results indicate that Cr-induced DNA interstrand crosslinks, and not DNA-Cr-GSH crosslinks, are the principal lesions responsible for blocking DNA replication. Moreover, the formation of DNA-Cr-GSH crosslinks may actually preclude the formation of the polymerase arresting lesions. (Mol Cell Biochem222:173-182, 2001)

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Authors and Affiliations

  1. Department of Pharmacology, The George Washington University Medical Center, Washington DC, USA

    Travis O’Brien

  2. Merck and Company, West Point, PA, USA

    Jian Xu

  3. Department of Pharmacology, The George Washington University Medical Center, 2300 Street N.W., Washington DC, 20037, USA

    Steven R. Patierno

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  1. Travis O’Brien
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  2. Jian Xu
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  3. Steven R. Patierno
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  1. Pathology and Physiology Research Branch, National Institute for Occupational Safety and Health, 1095 Willowdale Road, 26505, Morgantown, WV, USA

    Xianglin Shi , Vince Castranova  & Val Vallyathan ,  & 

  2. Occupational Safety and Health Administration, Washington DC, USA

    William G. Perry

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O’Brien, T., Xu, J., Patierno, S.R. (2001). Effects of Glutathione on Chromium-induced DNA Crosslinking and DNA Polymerase Arrest. In: Shi, X., Castranova, V., Vallyathan, V., Perry, W.G. (eds) Molecular Mechanisms of Metal Toxicity and Carcinogenesis. Developments in Molecular and Cellular Biochemistry, vol 34. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-0793-2_20

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