journal.pgen.1002407.pdf (1.08 MB)
The major roles of DNA polymerases epsilon and delta at the eukaryotic replication fork are evolutionarily conserved
journal contribution
posted on 2023-06-08, 12:53 authored by Izumi Miyabe, Thomas A Kunkel, Antony CarrAntony CarrCoordinated replication of eukaryotic genomes is intrinsically asymmetric, with continuous leading strand synthesis preceding discontinuous lagging strand synthesis. Here we provide two types of evidence indicating that, in fission yeast, these two biosynthetic tasks are performed by two different replicases. First, in Schizosaccharomyces pombe strains encoding a pold-L591M mutator allele, base substitutions in reporter genes placed in opposite orientations relative to a well-characterized replication origin are strand-specific and distributed in patterns implying that Pold is primarily involved in lagging strand replication. Second, in strains encoding a pole-M630F allele and lacking the ability to repair rNMPs in DNA due to a defect in RNase H2, rNMPs are selectively observed in nascent leading strand DNA. The latter observation demonstrates that abundant rNMP incorporation during replication can be tolerated and that they are normally removed in an RNase H2-dependent manner. This provides strong physical evidence that Pole is the primary leading strand replicase. Collectively, these data and earlier results in budding yeast indicate that the major roles of Pold and Pole at the eukaryotic replication fork are evolutionarily conserved.
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- Published
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- Published version
Journal
PLoS GeneticsISSN
1553-7390Publisher
Public Library of ScienceExternal DOI
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12Volume
7Article number
e1002407Department affiliated with
- Sussex Centre for Genome Damage Stability Publications
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- Yes
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- Yes
Legacy Posted Date
2012-10-29First Open Access (FOA) Date
2016-03-22First Compliant Deposit (FCD) Date
2017-03-06Usage metrics
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