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A noncatalytic function of the ligation complex during nonhomologous end joining
Nonhomologous end joining is the primary deoxyribonucleic acid (DNA) double-strand break repair pathway in multicellular eukaryotes. To initiate repair, Ku binds DNA ends and recruits the DNA-dependent protein kinase (DNA-PK) catalytic subunit (DNA-PKcs) forming the holoenzyme. Early end synapsis is...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3549972/ https://www.ncbi.nlm.nih.gov/pubmed/23337116 http://dx.doi.org/10.1083/jcb.201203128 |
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author | Cottarel, Jessica Frit, Philippe Bombarde, Oriane Salles, Bernard Négrel, Aurélie Bernard, Stéphanie Jeggo, Penny A. Lieber, Michael R. Modesti, Mauro Calsou, Patrick |
author_facet | Cottarel, Jessica Frit, Philippe Bombarde, Oriane Salles, Bernard Négrel, Aurélie Bernard, Stéphanie Jeggo, Penny A. Lieber, Michael R. Modesti, Mauro Calsou, Patrick |
author_sort | Cottarel, Jessica |
collection | PubMed |
description | Nonhomologous end joining is the primary deoxyribonucleic acid (DNA) double-strand break repair pathway in multicellular eukaryotes. To initiate repair, Ku binds DNA ends and recruits the DNA-dependent protein kinase (DNA-PK) catalytic subunit (DNA-PKcs) forming the holoenzyme. Early end synapsis is associated with kinase autophosphorylation. The XRCC4 (X4)–DNA Ligase IV (LIG4) complex (X4LIG4) executes the final ligation promoted by Cernunnos (Cer)–X4-like factor (XLF). In this paper, using a cell-free system that recapitulates end synapsis and DNA-PKcs autophosphorylation, we found a defect in both activities in human cell extracts lacking LIG4. LIG4 also stimulated the DNA-PKcs autophosphorylation in a reconstitution assay with purified components. We additionally uncovered a kinase autophosphorylation defect in LIG4-defective cells that was corrected by ectopic expression of catalytically dead LIG4. Finally, our data support a contribution of Cer-XLF to this unexpected early role of the ligation complex in end joining. We propose that productive end joining occurs by early formation of a supramolecular entity containing both DNA-PK and X4LIG4–Cer-XLF complexes on DNA ends. |
format | Online Article Text |
id | pubmed-3549972 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-35499722013-07-21 A noncatalytic function of the ligation complex during nonhomologous end joining Cottarel, Jessica Frit, Philippe Bombarde, Oriane Salles, Bernard Négrel, Aurélie Bernard, Stéphanie Jeggo, Penny A. Lieber, Michael R. Modesti, Mauro Calsou, Patrick J Cell Biol Research Articles Nonhomologous end joining is the primary deoxyribonucleic acid (DNA) double-strand break repair pathway in multicellular eukaryotes. To initiate repair, Ku binds DNA ends and recruits the DNA-dependent protein kinase (DNA-PK) catalytic subunit (DNA-PKcs) forming the holoenzyme. Early end synapsis is associated with kinase autophosphorylation. The XRCC4 (X4)–DNA Ligase IV (LIG4) complex (X4LIG4) executes the final ligation promoted by Cernunnos (Cer)–X4-like factor (XLF). In this paper, using a cell-free system that recapitulates end synapsis and DNA-PKcs autophosphorylation, we found a defect in both activities in human cell extracts lacking LIG4. LIG4 also stimulated the DNA-PKcs autophosphorylation in a reconstitution assay with purified components. We additionally uncovered a kinase autophosphorylation defect in LIG4-defective cells that was corrected by ectopic expression of catalytically dead LIG4. Finally, our data support a contribution of Cer-XLF to this unexpected early role of the ligation complex in end joining. We propose that productive end joining occurs by early formation of a supramolecular entity containing both DNA-PK and X4LIG4–Cer-XLF complexes on DNA ends. The Rockefeller University Press 2013-01-21 /pmc/articles/PMC3549972/ /pubmed/23337116 http://dx.doi.org/10.1083/jcb.201203128 Text en © 2013 Cottarel et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/). |
spellingShingle | Research Articles Cottarel, Jessica Frit, Philippe Bombarde, Oriane Salles, Bernard Négrel, Aurélie Bernard, Stéphanie Jeggo, Penny A. Lieber, Michael R. Modesti, Mauro Calsou, Patrick A noncatalytic function of the ligation complex during nonhomologous end joining |
title | A noncatalytic function of the ligation complex during nonhomologous end joining |
title_full | A noncatalytic function of the ligation complex during nonhomologous end joining |
title_fullStr | A noncatalytic function of the ligation complex during nonhomologous end joining |
title_full_unstemmed | A noncatalytic function of the ligation complex during nonhomologous end joining |
title_short | A noncatalytic function of the ligation complex during nonhomologous end joining |
title_sort | noncatalytic function of the ligation complex during nonhomologous end joining |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3549972/ https://www.ncbi.nlm.nih.gov/pubmed/23337116 http://dx.doi.org/10.1083/jcb.201203128 |
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