The impact of single-nucleotide polymorphisms of human apurinic/apyrimidinic endonuclease 1 on specific DNA binding and catalysis

Irina V. Alekseeva, Anastasiia T. Davletgildeeva, Olga V. Arkova, Nikita A. Kuznetsov, Olga S. Fedorova

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

Human apurinic/apyrimidinic (AP) endonuclease APE1 is a crucial enzyme of the base excision repair (BER) pathway, which is in charge of recognition and initiation of removal of AP-sites in DNA. It is known that some single-nucleotide polymorphism (SNP) variants of APE1 have a reduced activity as compared to wild-type APE1. It has been hypothesized that genetic variation in APE1 might be responsible for an increased risk of some types of cancer. In the present work, analysis of SNPs of the APE1 gene was performed to select the set of variants having substitutions of amino acid residues on the surface of the enzyme globule and in the DNA-binding site, thereby affecting protein–protein interactions or the catalytic reaction, respectively. For seven APE1 variants (R221C, N222H, R237A, G241R, M270T, R274Q, and P311S), conformational dynamics and catalytic activities were examined. The conformational changes in the molecules of APE1 variants and in a DNA substrate were recorded as fluorescence changes of Trp and 2-aminopurine residues, respectively, using the stopped-flow technique. The results made it possible to determine the kinetic mechanism underlying the interactions of the APE1 variants with DNA substrates, to calculate the rate constants of the elementary stages, and to identify the stages of the process affected by mutation.

Original languageEnglish
Pages (from-to)73-83
Number of pages11
JournalBiochimie
Volume163
DOIs
Publication statusPublished - 1 Aug 2019

Keywords

  • Abasic site
  • DNA repair
  • Fluorescence
  • Human apurinic/apyrimidinic endonuclease
  • Stopped-flow enzyme kinetics
  • RECOGNITION
  • ACTIVE-SITE
  • BIOLOGICAL CONSEQUENCES
  • Human apurinic/apyrimidinic endonudease
  • CONFORMATIONAL DYNAMICS
  • 2-AMINOPURINE
  • DAMAGE
  • BASE EXCISION-REPAIR
  • FLUORESCENCE
  • DIVALENT METAL-IONS
  • HUMAN AP ENDONUCLEASE
  • DNA/metabolism
  • Humans
  • Substrate Specificity
  • DNA-(Apurinic or Apyrimidinic Site) Lyase/chemistry
  • Models, Molecular
  • DNA Repair
  • Protein Conformation
  • Polymorphism, Single Nucleotide
  • DNA Damage
  • Kinetics
  • Mutation

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