Yeast DNA Polymerase ε Participates in Leading-Strand DNA Replication

Author:

Pursell Zachary F.12,Isoz Isabelle12,Lundström Else-Britt12,Johansson Erik12,Kunkel Thomas A.12

Affiliation:

1. Laboratory of Molecular Genetics and Laboratory of Structural Biology, National Institute of Environmental Health Sciences, NIH, DHHS, Research Triangle Park, NC 27709, USA.

2. Department of Medical Biochemistry and Biophysics, Umeå University, SE-901 87, Umeå, Sweden.

Abstract

Multiple DNA polymerases participate in replicating the leading and lagging strands of the eukaryotic nuclear genome. Although 50 years have passed since the first DNA polymerase was discovered, the identity of the major polymerase used for leading-strand replication is uncertain. We constructed a derivative of yeast DNA polymerase ϵ that retains high replication activity but has strongly reduced replication fidelity, particularly for thymine-deoxythymidine 5'-monophosphate (T-dTMP) but not adenine-deoxyadenosine 5'-monophosphate (A-dAMP) mismatches. Yeast strains with this DNA polymerase ϵ allele have elevated rates of T to A substitution mutations. The position and rate of these substitutions depend on the orientation of the mutational reporter and its location relative to origins of DNA replication and reveal a pattern indicating that DNA polymerase ϵ participates in leading-strand DNA replication.

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

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