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Homology and enzymatic requirements of microhomology-dependent alternative end joining

Nonhomologous DNA end joining (NHEJ) is one of the major double-strand break (DSB) repair pathways in higher eukaryotes. Recently, it has been shown that alternative NHEJ (A-NHEJ) occurs in the absence of classical NHEJ and is implicated in chromosomal translocations leading to cancer. In the presen...

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Autores principales: Sharma, S, Javadekar, S M, Pandey, M, Srivastava, M, Kumari, R, Raghavan, S C
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4385936/
https://www.ncbi.nlm.nih.gov/pubmed/25789972
http://dx.doi.org/10.1038/cddis.2015.58
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author Sharma, S
Javadekar, S M
Pandey, M
Srivastava, M
Kumari, R
Raghavan, S C
author_facet Sharma, S
Javadekar, S M
Pandey, M
Srivastava, M
Kumari, R
Raghavan, S C
author_sort Sharma, S
collection PubMed
description Nonhomologous DNA end joining (NHEJ) is one of the major double-strand break (DSB) repair pathways in higher eukaryotes. Recently, it has been shown that alternative NHEJ (A-NHEJ) occurs in the absence of classical NHEJ and is implicated in chromosomal translocations leading to cancer. In the present study, we have developed a novel biochemical assay system utilizing DSBs flanked by varying lengths of microhomology to study microhomology-mediated alternative end joining (MMEJ). We show that MMEJ can operate in normal cells, when microhomology is present, irrespective of occurrence of robust classical NHEJ. Length of the microhomology determines the efficiency of MMEJ, 5 nt being obligatory. Using this biochemical approach, we show that products obtained are due to MMEJ, which is dependent on MRE11, NBS1, LIGASE III, XRCC1, FEN1 and PARP1. Thus, we define the enzymatic machinery and microhomology requirements of alternative NHEJ using a well-defined biochemical system.
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spelling pubmed-43859362015-04-07 Homology and enzymatic requirements of microhomology-dependent alternative end joining Sharma, S Javadekar, S M Pandey, M Srivastava, M Kumari, R Raghavan, S C Cell Death Dis Original Article Nonhomologous DNA end joining (NHEJ) is one of the major double-strand break (DSB) repair pathways in higher eukaryotes. Recently, it has been shown that alternative NHEJ (A-NHEJ) occurs in the absence of classical NHEJ and is implicated in chromosomal translocations leading to cancer. In the present study, we have developed a novel biochemical assay system utilizing DSBs flanked by varying lengths of microhomology to study microhomology-mediated alternative end joining (MMEJ). We show that MMEJ can operate in normal cells, when microhomology is present, irrespective of occurrence of robust classical NHEJ. Length of the microhomology determines the efficiency of MMEJ, 5 nt being obligatory. Using this biochemical approach, we show that products obtained are due to MMEJ, which is dependent on MRE11, NBS1, LIGASE III, XRCC1, FEN1 and PARP1. Thus, we define the enzymatic machinery and microhomology requirements of alternative NHEJ using a well-defined biochemical system. Nature Publishing Group 2015-03 2015-03-19 /pmc/articles/PMC4385936/ /pubmed/25789972 http://dx.doi.org/10.1038/cddis.2015.58 Text en Copyright © 2015 Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Original Article
Sharma, S
Javadekar, S M
Pandey, M
Srivastava, M
Kumari, R
Raghavan, S C
Homology and enzymatic requirements of microhomology-dependent alternative end joining
title Homology and enzymatic requirements of microhomology-dependent alternative end joining
title_full Homology and enzymatic requirements of microhomology-dependent alternative end joining
title_fullStr Homology and enzymatic requirements of microhomology-dependent alternative end joining
title_full_unstemmed Homology and enzymatic requirements of microhomology-dependent alternative end joining
title_short Homology and enzymatic requirements of microhomology-dependent alternative end joining
title_sort homology and enzymatic requirements of microhomology-dependent alternative end joining
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4385936/
https://www.ncbi.nlm.nih.gov/pubmed/25789972
http://dx.doi.org/10.1038/cddis.2015.58
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