Repair of a minimal DNA double-strand break by NHEJ requires DNA-PKcs and is controlled by the ATM/ATR checkpoint
Open accessAbstract
Mammalian cells primarily rejoin DNA double-strand breaks (DSBs) by the non-homologous end-joining (NHEJ) pathway. The joining of the broken DNA ends appears directly without template and accuracy is ensured by the NHEJ factors that are under ATM/ATR regulated checkpoint control. In the current study we report the engineering of a mono-specific DNA damaging agent. This was used to study the molecular requirements for the repair of the least complex DSB in vivo. Single-chain PvuII restriction enzymes fused to protein delivery sequences transduce cells efficiently and induce blunt end DSBs in vivo. We demonstrate that beside XRCC4/LigaseIV and KU, the DNA-PK catalytic subunit (DNA-PKcs) is also essential for the joining of this low complex DSB in vivo. The appearance of blunt end 3'-hydroxyl and 5'-phosphate DNA DSBs induces a significantly higher frequency of anaphase bridges in cells that do not contain functional DNA-PKcs, suggesting an absolute requirement for DNA-PKcs in the control of chromosomal stability during end joining. Moreover, these minimal blunt end DSBs are sufficient to induce a p53 and ATM/ATR checkpoint function.
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ATM plays an important role in recruiting the Ku complex to hydrogen peroxide-damaged chromatin, supporting a role for ATM in double-strand break repair by nonhomologous end joining.
“Moreover, ATM plays an important role for the recruitment of Ku complex to hydrogen peroxide-damaged chromatin, further supporting the role of ATM in DSB repair by NHEJ [174, 175] (Fig. 6).”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [175]
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- Authors
- Kuhne, C.
- Affiliations
- AREA Science Park (IT)
- Corresponding author
- Kuhne, C.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2003-12-15
- Volume
- 31
- Issue
- 24
- Pages
- 7227-7237
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC291875
- OpenAlex
- W1965511479
- Cited by
- 36
- References
- 50
- Citations by year
- 2026: 2 · 2022: 2 · 2021: 3 · 2020: 1 · 2019: 1 · 2018: 1 · 2017: 3 · 2016: 1 · 2015: 1 · 2014: 3 · 2012: 1
- FWCI
- 0.69
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Nucleic Acids Research (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- DNA double-strand break repair, non-homologous end joining, DNA-PKcs, Ku heterodimer, anaphase bridges, chromosome stability, p53 activation, DNA end joining
- MeSH terms
- Ku Autoantigen, Animals, Cell Cycle, Cell Line, DNA, DNA Damage, DNA Repair, DNA Helicases, DNA-Binding Proteins, Hydrolysis, Rodentia, Deoxyribonucleases, Type II Site-Specific, Gene Products, tat, Tumor Suppressor Protein p53, Protein Serine-Threonine Kinases, Gene Deletion, Cell Cycle Proteins, Tumor Suppressor Proteins, Antigens, Nuclear, DNA-Activated Protein Kinase, Ataxia Telangiectasia Mutated Proteins
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