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PMID: 12065413 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

DNA damage-induced apoptosis requires the DNA-dependent protein kinase, and is mediated by the latent population of p53.

The EMBO journal ·Vol. 21 ·No. 12 ·2002-06-17 ·Pages 3000-8

Woo RA, Jack MT, Xu Y, Burma S, Chen DJ, Lee PW

Abstract

Mouse embryo fibroblasts (MEFs) expressing the adenovirus E1A protein undergo apoptosis upon exposure to ionizing radiation. We show here that immediately following gamma-irradiation, latent p53 formed a complex with the catalytic subunit of the DNA-dependent protein kinase (DNA-PK(CS)). The complex formation was DNase sensitive, suggesting that the proteins came together on the DNA, conceivably at strand breaks. This association was accompanied by phosphorylation of pre-existing, latent p53 at Ser18 (corresponding to Ser15 in human p53), which was not found in DNA-PK(CS)(-/-) cells. Most significantly, DNA damage-induced apoptosis was abolished in both DNA-PK(CS)(-/-) and p53(-/-) cells. In addition, blocking synthesis of inducible p53 by cycloheximide did not abrogate apoptosis, suggesting that the latent population of p53 is sufficient for executing the apoptotic program. Finally, E1A-expressing MEFs from a p53 "knock-in" mouse where Ser18 was mutated to an alanine had an attenuated apoptotic response, indicating that phosphorylation of this site by DNA-PK is a contributing factor for apoptosis.

Keywords
Non-programmatic
MeSH Terms
Animals Apoptosis/physiology Cell Fractionation Cells, Cultured Cycloheximide/pharmacology DNA Damage DNA-Activated Protein Kinase DNA-Binding Proteins Fibroblasts/drug effects,metabolism,radiation effects Gamma Rays Humans Mice Mice, Transgenic Nuclear Proteins Point Mutation Protein Binding Protein Serine-Threonine Kinases/genetics,metabolism Protein Synthesis Inhibitors/pharmacology Tumor Suppressor Protein p53/genetics,metabolism
Chemicals
DNA-Binding Proteins Nuclear Proteins Protein Synthesis Inhibitors Tumor Suppressor Protein p53 Cycloheximide DNA-Activated Protein Kinase PRKDC protein, human Prkdc protein, mouse Protein Serine-Threonine Kinases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Woo Richard A
Cancer Biology Research Group and Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada T2N 4N1.
Jack Melissa T
Xu Yang
Burma Sandeep
Chen David J
Lee Patrick W K
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdf307
Published
2002-06-17
Pages
3000-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC126062
Subset
IM
Grants
NCI NIH HHS · R01 CA050519 · United States
NCI NIH HHS · R37 CA050519 · United States
NIA NIH HHS · AG 17709 · United States
NIA NIH HHS · AG 50519 · United States
Corrections
ErratumIn
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External Links
PubMed source DOI: 10.1093/emboj/cdf307 PMC Full Text View journal details
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