title: Mechanisms that regulate localization of a DNA double-strand break to the nuclear periphery.
abstract: DNA double-strand breaks (DSBs) are among the most deleterious forms of DNA lesions in cells. Here we induced site-specific DSBs in yeast cells and monitored chromatin dynamics surrounding the DSB using Chromosome Conformation Capture (3C). We find that formation of a DSB within G1 cells is not sufficient to alter chromosome dynamics. However, DSBs formed within an asynchronous cell population result in large decreases in both intra- and interchromosomal interactions. Using live cell microscopy, we find that changes in chromosome dynamics correlate with relocalization of the DSB to the nuclear periphery. Sequestration to the periphery requires the nuclear envelope protein, Mps3p, and Mps3p-dependent tethering delays recombinational repair of a DSB and enhances gross chromosomal rearrangements. Furthermore, we show that components of the telomerase machinery are recruited to a DSB and that telomerase recruitment is required for its peripheral localization. Based on these findings, we propose that sequestration of unrepaired or slowly repaired DSBs to the nuclear periphery reflects a competition between alternative repair pathways.
authors.affiliation: Program in Molecular Medicine, Interdisciplinary Graduate Program, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
authors.forename: Pranav
authors.initials: P
authors.lastname: Oza
authors.forename: Sue L
authors.initials: SL
authors.lastname: Jaspersen
authors.forename: Adriana
authors.initials: A
authors.lastname: Miele
authors.forename: Job
authors.initials: J
authors.lastname: Dekker
authors.forename: Craig L
authors.initials: CL
authors.lastname: Peterson
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D015825:Chromosomes, Fungal / Q000235:genetics / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D004265:DNA Helicases / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D005821:Genetic Techniques; D008565:Membrane Proteins / Q000378:metabolism; D008853:Microscopy; D009687:Nuclear Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism; D034501:Telomere-Binding Proteins / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases
chemical_list: C452219:Cdc13 protein, S cerevisiae; D002843:Chromatin; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; C471174:Mps3 protein, S cerevisiae; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D034501:Telomere-Binding Proteins; D044767:Ubiquitin-Protein Ligases; D019098:Telomerase; C085463:PIF1 protein, S cerevisiae; D004265:DNA Helicases; C515573:Slx5 protein, S cerevisiae