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NEIL1 responds and binds to psoralen-induced DNA interstrand crosslinks

Daniel R McNeill1, Manikandan Paramasivam, Jakita Baldwin

  • 1Laboratory of Molecular Gerontology, Biomedical Research Center, NIA, National Institutes of Health, Baltimore, Maryland 21224, USA.

The Journal of Biological Chemistry
|March 20, 2013
PubMed

Insights

The DNA repair protein NEIL1 specifically binds to DNA interstrand crosslinks, potentially hindering their removal and impacting cellular repair pathways. This suggests a novel role for base excision repair (BER) proteins in crosslink processing.

Area of Science:

  • Molecular Biology
  • DNA Repair Mechanisms
  • Biochemistry

Background:

  • Base excision repair (BER) proteins are implicated in responding to DNA interstrand crosslinks (ICLs).
  • ICLs pose significant threats to cellular integrity by blocking replication and transcription, leading to genetic instability.
  • Understanding the specific roles of individual DNA repair proteins in ICL processing is crucial.

Purpose of the Study:

  • To investigate the role of the endonuclease VIII-like DNA glycosylase, NEIL1, in the cellular response to DNA interstrand crosslinks.
  • To elucidate the mechanisms by which NEIL1 recognizes and interacts with different types of DNA damage.
  • To determine the functional consequences of NEIL1 involvement in interstrand crosslink repair.

Main Methods:

  • Utilized fluorescently tagged fusion proteins and laser microirradiation coupled with confocal microscopy.
  • Observed NEIL1 accumulation at sites of oxidative DNA damage and trioxsalen-induced interstrand crosslinks.
  • Investigated the effects of antioxidants, NEIL1 variants, and XPC on NEIL1 recruitment.
  • Performed knockdown studies in glioblastoma cells and in vitro binding assays with synthetic DNA substrates.

Main Results:

  • NEIL1 accumulates at oxidative DNA damage and trioxsalen (psoralen)-induced interstrand crosslinks, but not angelicin monoadducts.
  • NEIL1 recruitment mechanisms differ for oxidative damage versus interstrand crosslinks, as shown by antioxidant treatment and NEIL1 variant studies.
  • NEIL1 recruitment to psoralen crosslinks is independent of the nucleotide excision repair factor XPC.
  • NEIL1 knockdown enhanced XPC recruitment, accelerated adduct removal, and decreased cellular sensitivity to trioxsalen plus UVA.
  • Purified NEIL1 protein stably bound to psoralen interstrand crosslink-containing DNA substrates in vitro, despite lacking enzymatic activity.

Conclusions:

  • NEIL1 specifically recognizes and binds to DNA interstrand crosslinks through distinct mechanisms.
  • NEIL1's interaction with interstrand crosslinks may obstruct efficient repair, potentially interfering with canonical repair pathways.
  • These findings reveal a novel, possibly inhibitory, role for NEIL1 in the cellular response to DNA interstrand crosslinks.
Keywords:
Base Excision RepairDNA CrosslinkDNA DamageDNA Damage ResponseDNA RepairNEIL1 GlycosylaseNonsynonymous PolymorphismProtein DNA-InteractionPsoralen

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