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PMID: 18093953 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Defects in XRCC4 and KU80 differentially affect the joining of distal nonhomologous ends.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 104 ·No. 52 ·2007-12-26 ·Pages 20902-7

Guirouilh-Barbat J, Rass E, Plo I, Bertrand P, Lopez BS

Abstract

XRCC4-null mice have a more severe phenotype than KU80-null mice. Here, we address whether this difference in phenotype is connected to nonhomologous end-joining (NHEJ). We used intrachromosomal substrates to monitor NHEJ of two distal double-strand breaks (DSBs) targeted by I-SceI, in living cells. In xrcc4-defective XR-1 cells, a residual but significant end-joining process exists, which primarily uses microhomologies distal from the DSB. However, NHEJ efficiency was strongly reduced in xrcc4-defective XR-1 cells versus complemented cells, contrasting with KU-deficient xrs6 cells, which showed levels of end-joining similar to those of complemented cells. Nevertheless, sequence analysis of the repair junctions indicated that the accuracy of end-joining was strongly affected in both xrcc4-deficient and KU-deficient cells. More specifically, these data showed that the KU80/XRCC4 pathway is conservative and not intrinsically error-prone but can accommodate non-fully complementary ends at the cost of limited mutagenesis.

MeSH Terms
Animals Antigens, Nuclear/genetics,physiology CD4-Positive T-Lymphocytes/metabolism CHO Cells Cricetinae Cricetulus DNA Damage DNA Repair DNA-Binding Proteins/genetics,physiology Flow Cytometry Gene Expression Regulation Genetic Complementation Test Ku Autoantigen Mice Mice, Transgenic Models, Genetic Phenotype
Chemicals
Antigens, Nuclear DNA-Binding Proteins XRCC4 protein, mouse Xrcc6 protein, mouse Ku Autoantigen
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Guirouilh-Barbat Josée
Commissariat à l'Energie Atomique, Unité Mixte de Recherche 217, Centre National de la Recherche Scientifique/Commissariat à l'Energie Atomique, Institut de Radiobiologie Cellulaire et Moléculaire, Fontenay aux Roses, France.
Rass Emilie
Plo Isabelle
Bertrand Pascale
Lopez Bernard S
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.0708541104
Published
2007-12-26
Epub
2007-00-18
Pages
20902-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2409239
Subset
IM
Corrections
CommentIn
PMID: 18180452
External Links
PubMed source DOI: 10.1073/pnas.0708541104 PMC Full Text View journal details
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