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Homology and enzymatic requirements of microhomology-dependent alternative end joining

Sharma, S and Javadekar, SM and Pandey, M and Srivastava, M and Kumari, R and Raghavan, SC (2015) Homology and enzymatic requirements of microhomology-dependent alternative end joining. In: CELL DEATH & DISEASE, 6 .

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Official URL: http://dx.doi.org/ 10.1038/cddis.2015.58

Abstract

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.

Item Type: Journal Article
Additional Information: Copy right for this article belongs to the NATURE PUBLISHING GROUP, MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND
Department/Centre: Division of Biological Sciences > Biochemistry
Depositing User: Id for Latest eprints
Date Deposited: 15 Jun 2015 07:31
Last Modified: 15 Jun 2015 07:31
URI: http://eprints.iisc.ac.in/id/eprint/51660

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