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PMID: 11134068 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

p53 binding protein 1 (53BP1) is an early participant in the cellular response to DNA double-strand breaks.

The Journal of cell biology ·Vol. 151 ·No. 7 ·2000-12-25 ·Pages 1381-90

Schultz LB, Chehab NH, Malikzay A, Halazonetis TD

Abstract

p53 binding protein 1 (53BP1), a protein proposed to function as a transcriptional coactivator of the p53 tumor suppressor, has BRCT domains with high homology to the Saccharomyces cerevisiae Rad9p DNA damage checkpoint protein. To examine whether 53BP1 has a role in the cellular response to DNA damage, we probed its intracellular localization by immunofluorescence. In untreated primary cells and U2OS osteosarcoma cells, 53BP1 exhibited diffuse nuclear staining; whereas, within 5-15 min after exposure to ionizing radiation (IR), 53BP1 localized at discreet nuclear foci. We propose that these foci represent sites of processing of DNA double-strand breaks (DSBs), because they were induced by IR and chemicals that cause DSBs, but not by ultraviolet light; their peak number approximated the number of DSBs induced by IR and decreased over time with kinetics that parallel the rate of DNA repair; and they colocalized with IR-induced Mre11/NBS and gamma-H2AX foci, which have been previously shown to localize at sites of DSBs. Formation of 53BP1 foci after irradiation was not dependent on ataxia-telangiectasia mutated (ATM), Nijmegen breakage syndrome (NBS1), or wild-type p53. Thus, the fast kinetics of 53BP1 focus formation after irradiation and the lack of dependency on ATM and NBS1 suggest that 53BP1 functions early in the cellular response to DNA DSBs.

MeSH Terms
Active Transport, Cell Nucleus/drug effects,radiation effects Amino Acid Sequence Androstadienes/pharmacology Antibodies, Monoclonal/immunology Antibody Specificity Ataxia Telangiectasia Mutated Proteins Caffeine/pharmacology Carrier Proteins/chemistry,immunology,metabolism Cell Cycle Proteins Cells, Cultured DNA/chemistry,genetics,metabolism,radiation effects DNA Damage/genetics,radiation effects DNA Repair/genetics DNA-Activated Protein Kinase DNA-Binding Proteins/metabolism Dose-Response Relationship, Drug Fibroblasts Fluorescent Antibody Technique Humans Intracellular Signaling Peptides and Proteins MRE11 Homologue Protein Molecular Sequence Data Mutation/genetics Nuclear Proteins/genetics,physiology Phosphoproteins Protein Serine-Threonine Kinases/genetics,metabolism,physiology Sequence Alignment Sequence Homology, Amino Acid Time Factors Tumor Cells, Cultured Tumor Suppressor Protein p53/metabolism Tumor Suppressor Proteins Tumor Suppressor p53-Binding Protein 1 Wortmannin Yeasts/chemistry
Chemicals
Androstadienes Antibodies, Monoclonal Carrier Proteins Cell Cycle Proteins DNA-Binding Proteins Intracellular Signaling Peptides and Proteins MRE11 protein, human Nuclear Proteins Phosphoproteins TP53BP1 protein, human Tumor Suppressor Protein p53 Tumor Suppressor Proteins Tumor Suppressor p53-Binding Protein 1 Caffeine DNA ATM protein, human Ataxia Telangiectasia Mutated Proteins DNA-Activated Protein Kinase PRKDC protein, human Protein Serine-Threonine Kinases MRE11 Homologue Protein Wortmannin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Schultz L B
Department of Molecular Genetics, The Wistar Institute, Philadelphia, Pennsylvania 19104, USA.
Chehab N H
Malikzay A
Halazonetis T D
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.151.7.1381
Published
2000-12-25
Pages
1381-90
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2150674
Subset
IM
Grants
NCI NIH HHS · CA76367 · United States
External Links
PubMed source DOI: 10.1083/jcb.151.7.1381 PMC Full Text View journal details
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