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Nucleic Acids Research 2006 34(5):1633-1645; doi:10.1093/nar/gkl064
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Published online 20 March 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Article

Genome wide distribution of illegitimate recombination events in Kluyveromyces lactis

Andreas Kegel, Paula Martinez, Sidney D. Carter and Stefan U. Åström*

Department of Developmental Biology, Wennergren Institute, Stockholm University SE-106 91 Stockholm, Sweden

*To whom correspondence should be addressed at Department of Developmental Biology, Wennergren Institute, Stockholm University, Arrhenius laboratories E3, SE-106 91 Stockholm, Sweden. Tel: 46 8 161566; Fax: 46 8 6126127; Email: stefan.astrom{at}devbio.su.se

Received December 21, 2005. Revised January 21, 2006. Accepted February 28, 2006.

Illegitimate recombination (IR) is the process by which two DNA molecules not sharing homology to each other are joined. In Kluyveromyces lactis, integration of heterologous DNA occurred very frequently therefore constituting an excellent model organism to study IR. IR was completely dependent on the nonhomologous end-joining (NHEJ) pathway for DNA double strand break (DSB) repair and we detected no other pathways capable of mediating IR. NHEJ was very versatile, capable of repairing both blunt and non-complementary ends efficiently. Mapping the locations of genomic IR-events revealed target site preferences, in which intergenic regions (IGRs) and ribosomal DNA were overrepresented six-fold compared to open reading frames (ORFs). The IGR-events occurred predominantly within transcriptional regulatory regions. In a rad52 mutant strain IR still preferentially occurred at IGRs, indicating that DSBs in ORFs were not primarily repaired by homologous recombination (HR). Introduction of ectopic DSBs resulted in the efficient targeting of IR to these sites, strongly suggesting that IR occurred at spontaneous mitotic DSBs. The targeting efficiency was equal when ectopic breaks were introduced in an ORF or an IGR. We propose that spontaneous DSBs arise more frequently in transcriptional regulatory regions and in rDNA and such DSBs can be mapped by analyzing IR target sites.


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S. D. Carter, S. Iyer, J. Xu, M. J. McEachern, and S. U. Astrom
The Role Of Nonhomologous End-Joining Components in Telomere Metabolism in Kluyveromyces lactis
Genetics, March 1, 2007; 175(3): 1035 - 1045.
[Abstract] [Full Text] [PDF]


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