TY - JOUR
T1 - Quantitative, Genome-Wide Analysis of Eukaryotic Replication Initiation and Termination
AU - McGuffee, Sean R.
AU - Smith, Duncan J.
AU - Whitehouse, Iestyn
N1 - Funding Information:
We thank Philip Zegerman (Gurdon Institute) for the SSDDCS strain and Ken Marians, Dirk Remus, Toshi Tsukiyama (Fred Hutchinson Cancer Research Center), and members of the Molecular Biology Program and Whitehouse Lab for discussions and comments on the manuscript. This work was supported by the National Institutes of Health (grant R01 GM102253) and an Alfred Bressler Scholars Endowment Award to I.W. D.J.S. is a Howard Hughes Medical Institute fellow of the Damon Runyon Cancer Research Foundation (DRG-#2046-10).
PY - 2013/4/11
Y1 - 2013/4/11
N2 - Many fundamental aspects of DNA replication, such as the exact locations where DNA synthesis is initiated and terminated, how frequently origins are used, and how fork progression is influenced by transcription, are poorly understood. Via the deep sequencing of Okazaki fragments, we comprehensively document replication fork directionality throughout the S. cerevisiae genome, which permits the systematic analysis of initiation, origin efficiency, fork progression, and termination. We show that leading-strand initiation preferentially occurs within a nucleosome-free region at replication origins. Using a strain in which late origins can be induced to fire early, we show that replication termination is a largely passive phenomenon that does not rely on cis-acting sequences or replication fork pausing. The replication profile is predominantly determined by the kinetics of origin firing, allowing us to reconstruct chromosome-wide timing profiles from an asynchronous culture.
AB - Many fundamental aspects of DNA replication, such as the exact locations where DNA synthesis is initiated and terminated, how frequently origins are used, and how fork progression is influenced by transcription, are poorly understood. Via the deep sequencing of Okazaki fragments, we comprehensively document replication fork directionality throughout the S. cerevisiae genome, which permits the systematic analysis of initiation, origin efficiency, fork progression, and termination. We show that leading-strand initiation preferentially occurs within a nucleosome-free region at replication origins. Using a strain in which late origins can be induced to fire early, we show that replication termination is a largely passive phenomenon that does not rely on cis-acting sequences or replication fork pausing. The replication profile is predominantly determined by the kinetics of origin firing, allowing us to reconstruct chromosome-wide timing profiles from an asynchronous culture.
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U2 - 10.1016/j.molcel.2013.03.004
DO - 10.1016/j.molcel.2013.03.004
M3 - Article
C2 - 23562327
AN - SCOPUS:84876073691
SN - 1097-2765
VL - 50
SP - 123
EP - 135
JO - Molecular Cell
JF - Molecular Cell
IS - 1
ER -