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Genome-wide identification and characterization of replication origins by deep sequencing

Author(s)
Xu, Jia; Yanagisawa, Yoshimi; Tsankov, Alexander M; Hart, Christopher; Aoki, Keita; Kommajosyula, Naveen; Steinmann, Kathleen E.; Bochicchio, James; Russ, Carsten; Regev, Aviv; Rando, Oliver J; Nusbaum, Chad; Niki, Hironori; Milos, Patrice M.; Weng, Zhiping; Rhind, Nicholas; Regev, Aviv; ... Show more Show less
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Abstract
Background DNA replication initiates at distinct origins in eukaryotic genomes, but the genomic features that define these sites are not well understood. Results We have taken a combined experimental and bioinformatic approach to identify and characterize origins of replication in three distantly related fission yeasts: Schizosaccharomyces pombe, Schizosaccharomyces octosporus and Schizosaccharomyces japonicus. Using single-molecule deep sequencing to construct amplification-free high-resolution replication profiles, we located origins and identified sequence motifs that predict origin function. We then mapped nucleosome occupancy by deep sequencing of mononucleosomal DNA from the corresponding species, finding that origins tend to occupy nucleosome-depleted regions. Conclusions The sequences that specify origins are evolutionarily plastic, with low complexity nucleosome-excluding sequences functioning in S. pombe and S. octosporus, and binding sites for trans-acting nucleosome-excluding proteins functioning in S. japonicus. Furthermore, chromosome-scale variation in replication timing is conserved independently of origin location and via a mechanism distinct from known heterochromatic effects on origin function. These results are consistent with a model in which origins are simply the nucleosome-depleted regions of the genome with the highest affinity for the origin recognition complex. This approach provides a general strategy for understanding the mechanisms that define DNA replication origins in eukaryotes.
Date issued
2012-04
URI
http://hdl.handle.net/1721.1/70949
Department
Massachusetts Institute of Technology. Department of Biology
Journal
Genome Biology
Publisher
Springer (Biomed Central Ltd.)
Citation
Xu, Jia et al. “Genome-wide Identification and Characterization of Replication Origins by Deep Sequencing.” Genome Biology 13.4 (2012): R27. Web. 25 May 2012.
Version: Final published version
ISSN
1465-6906
1474-7596

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