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Visualization of DNA Repair Proteins Interaction by Immunofluorescence

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Published on: June 26, 2020

Human DNA-PKcs promotes broken DNA-end structure independence during NHEJ.

Shingo Fujii¹, Mauro Modesti¹

¹Department of Genome Integrity, Cancer Research Center of Marseille, CNRS UMR7258, Inserm U1068, Institut Paoli-Calmettes, Aix Marseille University, 13273 Marseille, France.

|March 26, 2026

Summary

DNA-PKcs plays a significant role in DNA double-strand break repair via non-homologous end joining (NHEJ). This study reveals DNA-PKcs promotes efficient joining of both cohesive and blunt DNA ends, independent of their structure.

Keywords:

CP: genomics DNA double-strand break DNA-PKcs DNA-end structure NHEJ PAXX reconstitution

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Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

Non-homologous end joining (NHEJ) is the primary DNA double-strand break repair pathway in vertebrates.
The role of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) in NHEJ is debated, with conflicting evidence from different experimental approaches.

Purpose of the Study:

To resolve the discrepancy regarding the function of DNA-PKcs in NHEJ.
To investigate the precise role of DNA-PKcs in DNA end joining using reconstituted NHEJ systems.

Main Methods:

NHEJ reconstitution experiments were performed under optimized conditions.
The activity of DNA-PKcs in joining both cohesive and blunt DNA ends was assessed.

Main Results:

DNA-PKcs significantly promotes the joining of both cohesive and blunt DNA ends in an ATP-dependent manner.
The efficiency of joining cohesive and blunt ends was found to be indistinguishable in the presence of DNA-PKcs.

Conclusions:

DNA-PKcs facilitates a unique productive synaptic complex that drives NHEJ.
The function of DNA-PKcs in NHEJ is independent of the DNA end structure, resolving previous experimental discrepancies.