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The major roles of DNA polymerases epsilon and delta at the eukaryotic replication fork are evolutionarily conserved

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posted on 2023-06-08, 12:53 authored by Izumi Miyabe, Thomas A Kunkel, Antony CarrAntony Carr
Coordinated 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.

History

Publication status

  • Published

File Version

  • Published version

Journal

PLoS Genetics

ISSN

1553-7390

Publisher

Public Library of Science

Issue

12

Volume

7

Article number

e1002407

Department affiliated with

  • Sussex Centre for Genome Damage Stability Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2012-10-29

First Open Access (FOA) Date

2016-03-22

First Compliant Deposit (FCD) Date

2017-03-06

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