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PMID: 16061690 Published · ppublish English Journal Article Research Support, N.I.H., Extramural 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.

Independent and sequential recruitment of NHEJ and HR factors to DNA damage sites in mammalian cells.

The Journal of cell biology ·Vol. 170 ·No. 3 ·2005-08-01 ·Pages 341-7

Kim JS, Krasieva TB, Kurumizaka H, Chen DJ, Taylor AM, Yokomori K

Abstract

Damage recognition by repair/checkpoint factors is the critical first step of the DNA damage response. DNA double strand breaks (DSBs) activate checkpoint signaling and are repaired by nonhomologous end-joining (NHEJ) and homologous recombination (HR) pathways. However, in vivo kinetics of the individual factor responses and the mechanism of pathway choice are not well understood. We report cell cycle and time course analyses of checkpoint activation by ataxia-telangiectasia mutated and damage site recruitment of the repair factors in response to laser-induced DSBs. We found that MRN acts as a DNA damage marker, continuously localizing at unrepaired damage sites. Damage recognition by NHEJ factors precedes that of HR factors. HR factor recruitment is not influenced by NHEJ factor assembly and occurs throughout interphase. Damage site retention of NHEJ factors is transient, whereas HR factors persist at unrepaired lesions, revealing unique roles of the two pathways in mammalian cells.

MeSH Terms
Acid Anhydride Hydrolases Animals Antigens, Nuclear/genetics,metabolism Ataxia Telangiectasia Mutated Proteins Cell Cycle/physiology,radiation effects Cell Cycle Proteins/metabolism Cell Line Checkpoint Kinase 2 Chromosomal Proteins, Non-Histone DNA Damage/physiology DNA Repair/physiology DNA Repair Enzymes/metabolism DNA-Binding Proteins/genetics,metabolism Fungal Proteins/metabolism Histones/metabolism Humans Ku Autoantigen Lasers MRE11 Homologue Protein Mice Mice, Knockout Nuclear Proteins/metabolism Phosphorylation Protein Serine-Threonine Kinases/metabolism Protein Transport Recombination, Genetic Signal Transduction/physiology Tumor Suppressor Proteins/metabolism
Chemicals
Antigens, Nuclear Cell Cycle Proteins Chromosomal Proteins, Non-Histone DNA-Binding Proteins Fungal Proteins H2AX protein, human Histones MRE11 protein, human Mre11a protein, mouse NBN protein, human Nuclear Proteins Tumor Suppressor Proteins cohesins Checkpoint Kinase 2 ATM protein, human Ataxia Telangiectasia Mutated Proteins Atm protein, mouse CHEK2 protein, human Chek2 protein, mouse Protein Serine-Threonine Kinases MRE11 Homologue Protein Acid Anhydride Hydrolases Rad50 protein, human Xrcc6 protein, human Xrcc6 protein, mouse Ku Autoantigen DNA Repair Enzymes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kim Jong-Soo
Department of Biological Chemistry, School of Medicine, University of California, Irvine, CA 92697, USA.
Krasieva Tatiana B
Kurumizaka Hitoshi
Chen David J
Taylor A Malcolm R
Yokomori Kyoko
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200411083
Published
2005-08-01
Pages
341-7
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2171485
Subset
IM
Grants
NCI NIH HHS · CA100710 · United States
NCI NIH HHS · R37 CA050519 · United States
NCI NIH HHS · CA50519 · United States
NCI NIH HHS · R01 CA100710 · United States
NCI NIH HHS · R01 CA050519 · United States
External Links
PubMed source DOI: 10.1083/jcb.200411083 PMC Full Text View journal details
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