Cited by 36DNA Repair MechanismsEffects of Radiation ExposureDNA and Nucleic Acid Chemistry

Repair of a minimal DNA double-strand break by NHEJ requires DNA-PKcs and is controlled by the ATM/ATR checkpoint

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Kuhne, C.·2003·Nucleic Acids Research

Abstract

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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Authors
Kuhne, C.
Affiliations
AREA Science Park (IT)
Corresponding author
Kuhne, C.
Venue
Nucleic Acids Research
Published
2003-12-15
Volume
31
Issue
24
Pages
7227-7237
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
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
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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