Excision of Oxidatively Generated Guanine Lesions by Competing Base and Nucleotide Excision Repair Mechanisms in Human Cells

Vladimir Shafirovich, Konstantin Kropachev, Marina Kolbanovskiy, Nicholas Geacintov

Research output: Contribution to journalArticle

Abstract

The interchange between different repair mechanisms in human cells has long been a subject of interest. Here, we provide a direct demonstration that the oxidatively generated guanine lesions spiroiminodihydantoin (Sp) and 5-guanidinohydantoin (Gh) embedded in double-stranded DNA are substrates of both base excision repair (BER) and nucleotide excision repair (NER) mechanisms in intact human cells. Site-specifically modified, 32 P-internally labeled double-stranded DNA substrates were transfected into fibroblasts or HeLa cells, and the BER and/or NER mono- and dual incision products were quantitatively recovered after 2-8 h incubation periods and lysis of the cells. DNA duplexes bearing single benzo[a]pyrene-derived guanine adduct were employed as positive controls of NER. The NER activities, but not the BER activities, were abolished in XPA -/- cells, while the BER yields were strongly reduced in NEIL1 -/- cells. Co-transfecting different concentrations of analogous DNA sequences bearing the BER substrates 5-hydroxyuracil diminish the BER yields of Sp lesions and enhance the yields of NER products. These results are consistent with a model based on the local availability of BER and NER factors in human cells and their competitive binding to the same Sp or Gh BER/NER substrates.

Original languageEnglish (US)
JournalChemical research in toxicology
DOIs
StatePublished - Jan 1 2019

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Guanine
DNA Repair
Repair
Nucleotides
Cells
Bearings (structural)
Substrates
DNA
Competitive Binding
Benzo(a)pyrene
DNA sequences
Interchanges
HeLa Cells
Fibroblasts

ASJC Scopus subject areas

  • Toxicology

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Excision of Oxidatively Generated Guanine Lesions by Competing Base and Nucleotide Excision Repair Mechanisms in Human Cells. / Shafirovich, Vladimir; Kropachev, Konstantin; Kolbanovskiy, Marina; Geacintov, Nicholas.

In: Chemical research in toxicology, 01.01.2019.

Research output: Contribution to journalArticle

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