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Oxidative Damage Fine-Tunes G-Quadruplex Structures in Human Gene Promoters.

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Oxidative damage creates 8-oxoguanine (O8G), altering gene expression. This study reveals O8G stabilizes NEIL3 G-quadruplex structures, impacting gene regulation and cellular response to oxidative stress.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Structural Biology

Background:

  • Oxidative damage, specifically guanine to 8-oxoguanine (O8G) conversion, is linked to gene expression changes and genome instability.
  • The precise molecular mechanisms by which O8G influences these processes are not fully understood.

Purpose of the Study:

  • To investigate the structural impact of O8G modification on NEIL3 gene promoter G-quadruplexes (NEIL3-G4s).
  • To elucidate the role of O8G-induced structural changes in epigenetic regulation and cellular response to oxidative stress.

Main Methods:

  • Nuclear Magnetic Resonance (NMR) spectroscopy to determine the solution structures of native and O8G-modified NEIL3-G4s.
  • Functional assays to assess the impact of NEIL3-G4s on DNA polymerase activity and gene expression under oxidative stress.

Main Results:

  • NEIL3 promoter sequences form polymorphic parallel and hybrid G-quadruplex structures (NEIL3-G4s).
  • Site-specific O8G modification significantly reduces NEIL3-G4 polymorphism, stabilizing specific topologies like the (3+1) hybrid-1 G4.
  • A single G-to-O8G substitution induces a structural transition, demonstrating O8G's profound effect on G4-mediated epigenetic regulation.
  • Both native and O8G-modified NEIL3-G4s inhibit DNA polymerase activity.
  • NEIL3-G4 formation correlates with increased NEIL3 gene expression under oxidative stress.

Conclusions:

  • O8G modification induces specific structural rearrangements in NEIL3-G4s, impacting their function.
  • NEIL3-G4s act as sensors of oxidative damage and molecular switches for NEIL3 gene upregulation.
  • O8G-induced G4 structural plasticity is crucial for cellular responses to oxidative stress and gene expression regulation.