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Molecular and Cellular Biology, January 2000, p. 54-60, Vol. 20, No. 1
0270-7306/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Double-Strand Break in a Chromosomal LINE Element Can Be Repaired by Gene Conversion with Various Endogenous LINE Elements in Mouse Cells

Annie Tremblay,1 Maria Jasin,2 and Pierre Chartrand1,3,*

Molecular Biology Program1 and Department of Pathology and Cellular Biology,3 University of Montreal, Montreal, Quebec, Canada, and Cell Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center and Cornell University Graduate School of Medical Sciences, New York, New York2

Received 21 July 1999/Returned for modification 10 September 1999/Accepted 27 September 1999

A double-strand break (DSB) in the mammalian genome has been shown to be a very potent signal for the cell to activate repair processes. Two different types of repair have been identified in mammalian cells. Broken ends can be rejoined with or without loss or addition of DNA or, alternatively, a homologous template can be used to repair the break. For most genomic sequences the latter event would involve allelic sequences present on the sister chromatid or homologous chromosome. However, since more than 30% of our genome consists of repetitive sequences, these would have the option of using nonallelic sequences for homologous repair. This could have an impact on the evolution of these sequences and of the genome itself. We have designed an assay to look at the repair of DSBs in LINE-1 (L1) elements which number 105 copies distributed throughout the genome of all mammals. We introduced into the genome of mouse epithelial cells an L1 element with an I-SceI endonuclease site. We induced DSBs at the I-SceI site and determined their mechanism of repair. We found that in over 95% of cases, the DSBs were repaired by an end-joining process. However, in almost 1% of cases, we found strong evidence for repair involving gene conversion with various endogenous L1 elements, with some being used preferentially. In particular, the TF family and the L1Md-A2 subfamily, which are the most active in retrotransposition, appeared to be contributing the most in this process. The degree of homology did not seem to be a determining factor in the selection of the endogenous elements used for repair but may be based instead on accessibility. Considering their abundance and dispersion, gene conversion between repetitive elements may be occurring frequently enough to be playing a role in their evolution.

* Corresponding author. Mailing address: Centre Hospitalier de l'Université de Montréal (CHUM), Hôpital Notre-Dame, 1560 rue Sherbrooke Est, Montreal, Quebec, H2L 4M1, Canada. Phone: (514) 281-6000, ext. 6939. Fax: (514) 896-4689. E-mail: chartrap{at}magellan.umontreal.ca.

Molecular and Cellular Biology, January 2000, p. 54-60, Vol. 20, No. 1
0270-7306/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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