3' to 5' exonuclease activity located in the DNA polymerase delta subunit of Saccharomyces cerevisiae is required for accurate replication

In Saccharomyces cerevisiae, DNA polymerase delta (POLIII), the product of the CDC2 (POL3) gene, possesses, in its N-terminal half, the well conserved 3-domain 3' to 5' exonuclease site. Strains selectively mutagenized in this site display a mutator phenotype detected as a drastically incr...

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Bibliographic Details
Published in:The EMBO journal 1991-08, Vol.10 (8), p.2165-2170
Main Authors: Simon, M, Giot, L, Faye, G
Format: Article
Language:English
Subjects:
Amino Acid Sequence
amino acid sequences
Biological and medical sciences
DNA Polymerase III
DNA Replication
DNA, Fungal - biosynthesis
DNA-directed DNA polymerase
DNA-Directed DNA Polymerase - genetics
DNA-Directed DNA Polymerase - metabolism
exonuclease proofreading
Exonucleases - metabolism
Fundamental and applied biological sciences. Psychology
Genes, Fungal
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Mutagenesis, Site-Directed
Mutation
Plasmids
Replication
Restriction Mapping
Saccharomyces cerevisiae
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae - genetics
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Summary:In Saccharomyces cerevisiae, DNA polymerase delta (POLIII), the product of the CDC2 (POL3) gene, possesses, in its N-terminal half, the well conserved 3-domain 3' to 5' exonuclease site. Strains selectively mutagenized in this site display a mutator phenotype detected as a drastically increased spontaneous forward mutation rate to canavanine resistance or as an elevated reversion rate to lysine prototrophy. Assays on a partially purified extract of the mutant giving the largest mutator effect indicate that the 3' to 5' exonuclease activity is reduced below the detection limit whereas the DNA polymerizing activity has wild-type level. Therefore, our results provide experimental support for the hypothesis that the exonucleolytic proofreading activity associated with DNA polymerase delta resides on the DNA polymerase delta subunit and enhances the fidelity of DNA replication in east.
ISSN:0261-4189
1460-2075
DOI:10.1002/j.1460-2075.1991.tb07751.x