Related Experiment Video

Updated: Oct 31, 2025

A Simple, Rapid, and Quantitative Assay to Measure Repair of DNA-protein Crosslinks on Plasmids Transfected into Mammalian Cells
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A Simple, Rapid, and Quantitative Assay to Measure Repair of DNA-protein Crosslinks on Plasmids Transfected into Mammalian Cells

Published on: March 5, 2018

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Protein-oligonucleotide conjugates as model substrates for DNA-protein crosslink repair proteases

Hannah K Reinking1,2, Julian Stingele1,2

  • 1Department of Biochemistry, Ludwig Maximilians University, 81377 Munich, Germany.

STAR Protocols
|June 30, 2021
PubMed

Insights

DNA-protein crosslinks (DPCs) cause genome instability. Researchers developed novel DPC model substrates to study how Wss1/SPRTN proteases repair these damaging DNA lesions in vitro.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Covalent DNA-protein crosslinks (DPCs) are significant contributors to genomic instability.
  • The Wss1/SPRTN protease family is crucial for repairing DPCs.
  • A lack of appropriate in vitro substrates has limited the study of DPC repair enzyme specificity.

Purpose of the Study:

  • To create defined protein-oligonucleotide conjugates as model substrates for DPCs.
  • To facilitate the in vitro analysis of DPC protease activity and binding.

Main Methods:

  • Generation of artificial DPC model substrates using protein-oligonucleotide conjugates.
  • Utilizing these substrates in activity and binding assays for DPC proteases.

Main Results:

  • Successfully generated defined DPC model substrates.
  • These substrates enable detailed analysis of DPC protease function.

Conclusions:

  • The developed DPC model substrates are valuable tools for studying DNA repair mechanisms.
  • This advancement aids in understanding the specificity of Wss1/SPRTN proteases in DPC repair.
Keywords:
Molecular BiologyProtein Biochemistry

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