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Molecular and Cellular Biology, August 2007, p. 5261-5274, Vol. 27, No. 15
0270-7306/07/$08.00+0     doi:10.1128/MCB.01852-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Interchromosomal Crossover in Human Cells Is Associated with Long Gene Conversion Tracts{triangledown}

Efrem A. H. Neuwirth,1 Masamitsu Honma,2 and Andrew J. Grosovsky1*

Department of Cell Biology and Neuroscience and Environmental Toxicology Graduate Program, University of California, Riverside, California 92521,1 Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya, Tokyo, Japan2

Received 30 September 2006/ Returned for modification 2 November 2006/ Accepted 23 April 2007

Crossovers have rarely been observed in specific association with interchromosomal gene conversion in mammalian cells. In this investigation two isogenic human B-lymphoblastoid cell lines, TI-112 and TSCER2, were used to select for I-SceI-induced gene conversions that restored function at the selectable thymidine kinase locus. Additionally, a haplotype linkage analysis methodology enabled the rigorous detection of all crossover-associated convertants, whether or not they exhibited loss of heterozygosity. This methodology also permitted characterization of conversion tract length and structure. In TI-112, gene conversion tracts were required to be complex in tract structure and at least 7.0 kb in order to be selectable. The results demonstrated that 85% (39/46) of TI-112 convertants extended more than 11.2 kb and 48% also exhibited a crossover, suggesting a mechanistic link between long tracts and crossover. In contrast, continuous tracts as short as 98 bp are selectable in TSCER2, although selectable gene conversion tracts could include a wide range of lengths. Indeed, only 16% (14/95) of TSCER2 convertants were crossover associated, further suggesting a link between long tracts and crossover. Overall, these results demonstrate that gene conversion tracts can be long in human cells and that crossovers are observable when long tracts are recoverable.

* Corresponding author. Mailing address: University of California, Department of Cell Biology and Neuroscience, 2211 Biological Sciences Building, Riverside, CA 92521. Phone: (951) 827-3193. Fax: (951) 827-3087. E-mail: grosovsky{at}ucr.edu

{triangledown} Published ahead of print on 21 May 2007.

Molecular and Cellular Biology, August 2007, p. 5261-5274, Vol. 27, No. 15
0270-7306/07/$08.00+0     doi:10.1128/MCB.01852-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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