Regulatory regions responsive to oxidative stress in the promoter of the human DNA glycosylase gene NEIL2

Carla J. Kinslow, Randa A. El-Zein, Catherine M. Rondelli, Courtney E. Hill, Jeffrey K. Wickliffe, Sherif Abdel-Rahman

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Reactive oxygen species (ROS) generated endogenously or from exogenous sources produce mutagenic DNA lesions. If not repaired, these lesions could lead to genomic instability and, potentially, to cancer development. NEIL2 (EC 4.2.99.18), a mammalian base excision repair (BER) protein and ortholog of the bacterial Fpg/Nei, excises oxidized DNA lesions from bubble or single-stranded structures, suggesting its involvement in transcription-coupled DNA repair. Perturbation in NEIL2 expression may, therefore, significantly impact BER capacity and promote genomic instability. To characterize the genetic and environmental factors regulating NEIL2 gene expression, we mapped the human NEIL2 transcriptional start site and partially characterized the promoter region of the gene using a luciferase reporter assay. We identified a strong positive regulatory region from nucleotide -206 to +90 and found that expression from this region was contingent on its being isolated from an adjacent strong negative regulatory region located downstream (+49 to +710 bp), suggesting that NEIL2 transcription is influenced by both these regions. We also found that oxidative stress, induced by glucose oxidase treatment, reduced the positive regulatory region expression levels, suggesting that ROS may play a significant role in regulating NEIL2 transcription. In an initial attempt to characterize the underlying mechanisms, we used in silico analysis to identify putative cis-acting binding sites for ROS-responsive transcription factors within this region and then used site-directed mutagenesis to investigate their role. A single-base change in the region encompassing nucleotides -206 to +90 abolished the effect of oxidative stress that was observed in the absence of the mutation. Our study is the first to provide an initial partial characterization of the NEIL2 promoter and opens the door for future research aimed at understanding the role of genetic and environmental factors in regulating NEIL2 expression.

Original languageEnglish (US)
Pages (from-to)171-177
Number of pages7
JournalMutagenesis
Volume25
Issue number2
DOIs
StatePublished - Mar 2010

Fingerprint

DNA Glycosylases
Oxidative stress
Nucleic Acid Regulatory Sequences
Transcription
DNA Repair
Reactive Oxygen Species
Oxidative Stress
Repair
Genes
Genomic Instability
DNA
Nucleotides
Glucose Oxidase
Mutagenesis
Bacterial Proteins
Site-Directed Mutagenesis
Luciferases
Genetic Promoter Regions
Gene expression
Computer Simulation

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Regulatory regions responsive to oxidative stress in the promoter of the human DNA glycosylase gene NEIL2. / Kinslow, Carla J.; El-Zein, Randa A.; Rondelli, Catherine M.; Hill, Courtney E.; Wickliffe, Jeffrey K.; Abdel-Rahman, Sherif.

In: Mutagenesis, Vol. 25, No. 2, 03.2010, p. 171-177.

Research output: Contribution to journalArticle

Kinslow, Carla J. ; El-Zein, Randa A. ; Rondelli, Catherine M. ; Hill, Courtney E. ; Wickliffe, Jeffrey K. ; Abdel-Rahman, Sherif. / Regulatory regions responsive to oxidative stress in the promoter of the human DNA glycosylase gene NEIL2. In: Mutagenesis. 2010 ; Vol. 25, No. 2. pp. 171-177.
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