Cited by 33DNA Repair MechanismsDNA and Nucleic Acid ChemistryEffects of Radiation Exposure

A mechanism for DNA-PK activation requiring unique contributions from each strand of a DNA terminus and implications for microhomology-mediated nonhomologous DNA end joining

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Pawelczak, Katherine S., Turchi, John J.·2008·Nucleic Acids Research

Abstract

DNA-dependent protein kinase (DNA-PK) is an essential component of the nonhomologous end joining pathway (NHEJ), responsible for the repair of DNA double-strand breaks. Ku binds a DSB and recruits the catalytic subunit, DNA-PKcs, where it is activated once the kinase is bound to the DSB. The precise mechanism by which DNA activates DNA-PK remains unknown. We have investigated the effect of DNA structure on DNA-PK activation and results demonstrate that in Ku-dependent DNA-PKcs reactions, DNA-PK activation with DNA effectors containing two unannealed ends was identical to activation observed with fully duplex DNA effectors of the same length. The presence of a 6-base single-stranded extension resulted in decreased activation compared to the fully duplex DNA. DNA-PK activation using DNA effectors with compatible termini displayed increased activity compared to effectors with noncompatible termini. A strand orientation preference was observed in these reactions and suggests a model where the 3' strand of the terminus is responsible for annealing and the 5' strand is involved in activation of DNA-PK. These results demonstrate the influence of DNA structure and orientation on DNA-PK activation and provide a molecular mechanism of activation resulting from compatible termini, an essential step in microhomology-mediated NHEJ.

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  1. [32]Hammarsten O DeFazio LG Chu G Activation of DNA-dependent protein kinase by single-stranded DNA ends J. Biol. Chem 2000 275 1541 1550 10636842

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Authors
Pawelczak, K. S., & Turchi, J. J.
Affiliations
Indiana University Indianapolis (US)
Corresponding author
Turchi, John J.
Venue
Nucleic Acids Research
Published
2008-05-31
Volume
36
Issue
12
Pages
4022-4031
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
OpenAlex
W2009896547
Cited by
33
References
33
Citations by year
2026: 1 · 2025: 1 · 2024: 2 · 2022: 2 · 2021: 2 · 2020: 2 · 2017: 4 · 2016: 1 · 2015: 3 · 2014: 2 · 2013: 3 · 2012: 1
FWCI
1.10
Type
journal article
Language
en
Open access
Locations
  • Nucleic Acids Research (publishedVersion, cc-by-nc)
  • PubMed Central (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
  • IUScholarWorks (Indiana University) (submittedVersion, cc-by-nc)
Funders
  • Flight Attendant Medical Research Institute
  • National Institute for Health and Care Research
  • National Institutes of Health
  • National Cancer Institute (CA82741, R01CA082741)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Effects of Radiation Exposure
Keywords
DNA-dependent protein kinase, Ku protein, non-homologous end joining, DNA double-strand break repair, single-stranded DNA, strand orientation, DNA annealing
MeSH terms
DNA, DNA Repair, Enzyme Activation, Models, Biological, Phosphorylation, Sequence Homology, Nucleic Acid, DNA-Activated Protein Kinase

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