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Nat Struct Mol Biol. 2015 Mar;22(3):192-198. doi: 10.1038/nsmb.2962. Epub 2015 Feb 9.

A global profile of replicative polymerase usage.

Author information

1
Genome Damage and Stability Centre, University of Sussex, Brighton, UK.
2
Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.
3
Centre for Translational Omics, University College London Institute of Child Health, London, UK.
4
School of Life Sciences, University of Nottingham Queens Medical Centre, Nottingham, UK.
#
Contributed equally

Abstract

Three eukaryotic DNA polymerases are essential for genome replication. Polymerase (Pol) α-primase initiates each synthesis event and is rapidly replaced by processive DNA polymerases: Polɛ replicates the leading strand, whereas Polδ performs lagging-strand synthesis. However, it is not known whether this division of labor is maintained across the whole genome or how uniform it is within single replicons. Using Schizosaccharomyces pombe, we have developed a polymerase usage sequencing (Pu-seq) strategy to map polymerase usage genome wide. Pu-seq provides direct replication-origin location and efficiency data and indirect estimates of replication timing. We confirm that the division of labor is broadly maintained across an entire genome. However, our data suggest a subtle variability in the usage of the two polymerases within individual replicons. We propose that this results from occasional leading-strand initiation by Polδ followed by exchange for Polɛ.

PMID:
25664722
PMCID:
PMC4789492
DOI:
10.1038/nsmb.2962
[Indexed for MEDLINE]
Free PMC Article

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