Документ взят из кэша поисковой машины. Адрес оригинального документа : http://mccmb.belozersky.msu.ru/2013/abstracts/abstracts/116.pdf Дата изменения: Mon Mar 18 14:39:00 2013 Дата индексирования: Thu Feb 27 20:54:09 2014 Кодировка:

Landscape of double-stranded DNA breaks in human genome and its correlation with sequence motifs and DNA bendability


We have developed a genome-
wide approach to map DNA double-
strand breaks (DSBs) at nucleotide

resolution by a method we termed BLESS(direct in situ breaks labeling, enrichment on streptavidin and next-
generation sequencing). We validated and tested BLESSusing human and mouse cells and different DSBs-
inducing agents and sequencing platforms. Our method is suitable for genome-
wide mapping of DSBs in various cells and experimental conditions, with a specificity and resolution unachievable by current techniques. We characterized the genomic landscape of sensitivity to replication stress in human cells, and we identified >2,000 nonuniformly distributed aphidicolin-
sensitive regions (ASRs) overrepresented in genes and enriched in satellite repeats. ASRs were also enriched in regions rearranged in human cancers, with many cancer-
associated genes exhibiting high sensitivity to replication stress. This exciting result contributes to understanding why some genes are more often mutated in cancers. Our data also shows that double-
stranded breaks occur more often in transcribed regions, thus supporting the hypothesis that collisions between transcription and replication machineries contribute substantially to DNA double-
breaks formation. We are currently model in silico movements and collisions of these complexes and will verify the computational predictions experimentally.




Reference: N. Crosetto, A. Mitra, M. J. Silva, M. Bienko, N. Dojer, Q. Wang, E. Karaca, R. Chiarle, M. Skrzypczak, K. Ginalski, P. Pasero, M. Rowicka (a corresponding author) & Ivan Dikic: Nucleotide-resolution DNA double-strand break mapping by next-generation sequencing, Nature Methods, in press.