Small-Molecule Inhibitor Screen for DNA Repair Proteins

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• The DNA damage response (DDR) and DNA repair pathways are crucial for maintaining genomic stability.
• Targeting proteins involved in DDR and repair offers therapeutic potential, but requires novel screening methods.
• Many DDR proteins interact directly with DNA, presenting a viable target class for small molecule intervention.

Purpose of the Study:

• To develop a high-throughput screening (HTS) assay for identifying modulators of protein-nucleic acid interactions.
• To enable the discovery of small molecules that can perturb DNA-protein interactions relevant to DDR and DNA repair.

Main Methods:

• A solution-based equilibrium binding fluorescence polarization (FP) assay was developed.
• The assay is designed for high-throughput screening of diverse protein-nucleic acid interactions.
• The FP assay measures the binding of small molecules to DNA-protein complexes.

Main Results:

• The developed FP assay is sensitive and stable for detecting binding events.
• The assay successfully identified small molecules capable of blocking DNA-protein interactions.
• This methodology is applicable to a broad range of protein-nucleic acid interactions.

Conclusions:

• A novel, high-throughput fluorescence polarization assay enables the screening of DNA-protein interactions.
• This assay expands the scope of targetable proteins in DDR and DNA repair beyond kinases.
• The method facilitates the identification of small molecules for perturbing essential protein-nucleic acid interactions.