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Nucleic Acids Research Advance Access published online on October 25, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm888
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© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-com

Molecular Biology

The ERCC1/XPF endonuclease is required for efficient single-strand annealing and gene conversion in mammalian cells

Ali Z. Al-Minawi1,2, Nasrollah Saleh-Gohari1,3 and Thomas Helleday1,2,4,*

1The Institute for Cancer Studies, University of Sheffield, Medical School, Beech Hill Road, Sheffield S10 2RX, UK, 2Department of Genetics Microbiology and Toxicology, Arrhenius Laboratory, Stockholm University, S-106 91 Stockholm, Sweden, 3The Afzalipour Hospital, Kerman University of Medical Science, Kerman, Iran and 4Radiation Oncology and Biology, University of Oxford, Oxford OX3 7LJ, UK

* To whom correspondence should be addressed. Tel: +44 1865 225 851; Fax: +44 1865 857 127; Email: thomas.helleday{at}rob.ox.ac.uk

Received September 6, 2007. Revised October 2, 2007. Accepted October 2, 2007.

The mammalian ERCC1-XPF endonuclease has a suggested role in the repair of DNA double-strand breaks (DSB) by single-strand annealing (SSA). Here, we investigated the role of ERCC1 in homologous recombination in mammalian cells, and confirm a role of ERCC1 in SSA. Interestingly, we also report an unexpected role for ERCC1 in gene conversion. This provides support that gene conversion in mammalian somatic cells is carried out through synthesis-dependent strand annealing, rather than through a double Holliday Junction mechanism. Moreover, we find low frequencies of SSA and gene conversion in G1-arrested cells, suggesting that SSA is not a frequent DSB repair pathway in G1-arrested mammalian cells, even in the presence of perfect repeats. Furthermore, we find that SSA is not influenced by inhibition of CDK2 (using Roscovitine), ATM (using Caffeine and KU55933), Chk1 (using CEP-3891) or DNA-PK (using NU7026).


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