Base and nucleotide excision repair of oxidatively generated guanine lesions in DNA

Vladimir Shafirovich, Konstantin Kropachev, Thomas Anderson, Zhi Liu, Marina Kolbanovskiy, Brooke D. Martin, Kent Sugden, Yoonjung Shim, Xuejing Chen, Jung Hyun Min, Nicholas E. Geacintov

Research output: Contribution to journalArticle

Abstract

The well known biomarker of oxidative stress, 8-oxo-7, 8-dihydroguanine, is more susceptible to further oxidation than the parent guanine base and can be oxidatively transformed to the genotoxic spiroiminodihydantoin (Sp) and 5-guanidinohydantoin (Gh) lesions. Incubation of 135-mer duplexes with single Sp or Gh lesions in human cell extracts yields a characteristic nucleotide excision repair (NER)-induced ladder of short dual incision oligonucleotide fragments in addition to base excision repair (BER) incision products. The ladders were not observed when NER was inhibited either by mouse monoclonal antibody (5F12) to human XPA or in XPC-/- fibroblast cell extracts. However, normal NER activity appeared when the XPC-/- cell extracts were complemented with XPC-RAD23B proteins. The Sp and Gh lesions are excellent substrates of both BER and NER. In contrast, 5-guanidino-4-nitroimidazole, a product of the oxidation of guanine in DNA by peroxynitrite, is an excellent substrate of BER only. In the case of mouse embryonic fibroblasts, BER of the Sp lesion is strongly reduced in NEIL1-/- relative to NEIL1+/+ extracts. In summary, in human cell extracts, BER and NER activities co-exist and excise Gh and Sp DNA lesions, suggesting that the relative NER/BER product ratios may depend on competitive BER and NER protein binding to these lesions.

Original languageEnglish (US)
Pages (from-to)5309-5319
Number of pages11
JournalJournal of Biological Chemistry
Volume291
Issue number10
DOIs
StatePublished - Mar 4 2016

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Base and nucleotide excision repair of oxidatively generated guanine lesions in DNA'. Together they form a unique fingerprint.

  • Cite this

    Shafirovich, V., Kropachev, K., Anderson, T., Liu, Z., Kolbanovskiy, M., Martin, B. D., Sugden, K., Shim, Y., Chen, X., Min, J. H., & Geacintov, N. E. (2016). Base and nucleotide excision repair of oxidatively generated guanine lesions in DNA. Journal of Biological Chemistry, 291(10), 5309-5319. https://doi.org/10.1074/jbc.M115.693218