rapid analysis of saccharomyces cerevisiae genome rearrangements by multiplex ligation–dependent probe amplification快速分析重组酿酒酵母基因组的多路复用ligation-dependent探测器放大.pdfVIP

Rapid Analysis of Saccharomyces cerevisiae Genome Rearrangements by Multiplex Ligation–Dependent Probe Amplification Jason E. Chan1,2,3, Richard D. Kolodner2,3* 1 Bioinformatics and Systems Biology Graduate Program, University of California San Diego, La Jolla, California, United States of America, 2 Ludwig Institute for Cancer Research, Cancer Center and Departments of Medicine and Cellular and Molecular Medicine, Moores–UCSD Cancer Center, School of Medicine, University of California San Diego, La Jolla, California, United States of America, 3 Institute of Genomic Medicine, School of Medicine, University of California San Diego, La Jolla, California, United States of America Abstract Aneuploidy and gross chromosomal rearrangements (GCRs) can lead to genetic diseases and the development of cancer. We previously demonstrated that introduction of the repetitive retrotransposon Ty912 onto a nonessential chromosome arm of Saccharomyces cerevisiae led to increased genome instability predominantly due to increased rates of formation of monocentric nonreciprocal translocations. In this study, we adapted Multiplex Ligation–dependent Probe Amplification (MLPA) to analyze a large numbers of these GCRs. Using MLPA, we found that the distribution of translocations induced by the presence of Ty912 in a wild-type strain was nonrandom and that the majority of these translocations were mediated by only six translocation targets on four different chromosomes, even though there were 254 potential Ty-related translocation targets in the S. cerevisiae genome. While the majority of Ty912-mediated translocations resulted from RAD52-dependent recombination, we observed a number of nonreciprocal translocations mediated by RAD52-independent recombination between Ty1 elements. The formation of these RAD52-independent translocations did not require the Rad51 or Rad59 homologo