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PMID: 12065431 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Synapsis of DNA ends by DNA-dependent protein kinase.

The EMBO journal ·Vol. 21 ·No. 12 ·2002-06-17 ·Pages 3192-200

DeFazio LG, Stansel RM, Griffith JD, Chu G

Abstract

The catalytic subunit of DNA-dependent protein kinase (DNA-PK(CS)) is required for a non-homologous end-joining pathway that repairs DNA double-strand breaks produced by ionizing radiation or V(D)J recombination; however, its role in this pathway has remained obscure. Using a neutravidin pull-down assay, we found that DNA-PK(CS) mediates formation of a synaptic complex containing two DNA molecules. Furthermore, kinase activity was cooperative with respect to DNA concentration, suggesting that activation of the kinase occurs only after DNA synapsis. Electron microscopy revealed complexes of two DNA ends brought together by two DNA-PK(CS) molecules. Our results suggest that DNA-PK(CS) brings DNA ends together and then undergoes activation of its kinase, presumably to regulate subsequent steps for processing and ligation of the ends.

MeSH Terms
Antigens, Nuclear Chromosome Pairing/physiology DNA/genetics,metabolism,ultrastructure DNA Damage DNA Helicases DNA Repair DNA-Activated Protein Kinase DNA-Binding Proteins/metabolism Enzyme Activation HeLa Cells Humans Ku Autoantigen Models, Genetic Nuclear Proteins/metabolism Nucleic Acid Conformation Protein Serine-Threonine Kinases/metabolism
Chemicals
Antigens, Nuclear DNA-Binding Proteins Nuclear Proteins DNA DNA-Activated Protein Kinase PRKDC protein, human Protein Serine-Threonine Kinases DNA Helicases XRCC5 protein, human Xrcc6 protein, human Ku Autoantigen
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
DeFazio Lisa G
Departments of Medicine and Biochemistry, Stanford University, Stanford, CA 94305, USA.
Stansel Rachel M
Griffith Jack D
Chu Gilbert
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdf299
Published
2002-06-17
Pages
3192-200
Language
English
Region
England
NLM ID
8208664
PMCID
PMC126055
Subset
IM
Grants
NCI NIH HHS · CA 70343 · United States
NIGMS NIH HHS · R01 GM 58120 · United States
NIGMS NIH HHS · R01 GM 31819 · United States
NIGMS NIH HHS · R01 GM031819 · United States
NIGMS NIH HHS · R01 GM058120 · United States
External Links
PubMed source DOI: 10.1093/emboj/cdf299 PMC Full Text View journal details
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