Effect of oxidative DNA damage in promoter elements on transcription factor binding

Rita Ghosh, David L. Mitchell

Research output: Contribution to journalArticle

103 Citations (Scopus)

Abstract

Reactive oxygen species produced by endogenous metabolic activity and exposure to a multitude of exogenous agents impact cells in a variety of ways. The DNA base damage 8-oxodeoxyguanosine (8-oxodG) is a prominent indicator of oxidative stress and has been well-characterized as a premutagenic lesion in mammalian cells and putative initiator of the carcinogenic process. Commensurate with the recent interest in epigenetic pathways of cancer causation we investigated how 8-oxodG alters the interaction between cis elements located on gene promoters and sequence-specific DNA binding proteins associated with these promoters. Consensus binding sequences for the transcription factors AP-1, NF-κB and Sp1 were modified site-specifically at guanine residues and electrophoretic mobility shift assays were performed to assess DNA-protein interactions. Our results indicate that whereas a single 8-oxodG was sufficient to inhibit transcription factor binding to AP-1 and Sp1 sequences it had no effect on binding to NF-κB, regardless of its position. We conclude from these data that minor alterations in base composition at a crucial position within some, but not all, promoter elements have the ability to disrupt transcription factor binding. The lack of inhibition by damaged NF-κB sequences suggests that DNA-protein contact sites may not be as determinative for stable p50 binding to this promoter as other, as yet undefined, structural parameters.

Original languageEnglish (US)
Pages (from-to)3213-3218
Number of pages6
JournalNucleic Acids Research
Volume27
Issue number15
DOIs
StatePublished - Aug 1 1999
Externally publishedYes

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Transcription Factor AP-1
DNA Damage
Transcription Factors
Consensus Sequence
Base Composition
DNA-Binding Proteins
Guanine
Electrophoretic Mobility Shift Assay
Epigenomics
Causality
Reactive Oxygen Species
Proteins
Oxidative Stress
DNA
Genes
Neoplasms

ASJC Scopus subject areas

  • Genetics

Cite this

Effect of oxidative DNA damage in promoter elements on transcription factor binding. / Ghosh, Rita; Mitchell, David L.

In: Nucleic Acids Research, Vol. 27, No. 15, 01.08.1999, p. 3213-3218.

Research output: Contribution to journalArticle

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