Cargando…

Evidence for interplay among yeast replicative DNA polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations

BACKGROUND: DNA polymerase ε (Pol ε) is essential for S-phase replication, DNA damage repair and checkpoint control in yeast. A pol2-Y831A mutation leading to a tyrosine to alanine change in the Pol ε active site does not cause growth defects and confers a mutator phenotype that is normally subtle b...

Descripción completa

Detalles Bibliográficos
Autores principales: Pavlov, Youri I, Maki, Satoko, Maki, Hisaji, Kunkel, Thomas A
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2004
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC434536/
https://www.ncbi.nlm.nih.gov/pubmed/15163346
http://dx.doi.org/10.1186/1741-7007-2-11
_version_ 1782121526994141184
author Pavlov, Youri I
Maki, Satoko
Maki, Hisaji
Kunkel, Thomas A
author_facet Pavlov, Youri I
Maki, Satoko
Maki, Hisaji
Kunkel, Thomas A
author_sort Pavlov, Youri I
collection PubMed
description BACKGROUND: DNA polymerase ε (Pol ε) is essential for S-phase replication, DNA damage repair and checkpoint control in yeast. A pol2-Y831A mutation leading to a tyrosine to alanine change in the Pol ε active site does not cause growth defects and confers a mutator phenotype that is normally subtle but strong in a mismatch repair-deficient strain. Here we investigate the mechanism responsible for the mutator effect. RESULTS: Purified four-subunit Y831A Pol ε turns over more deoxynucleoside triphosphates to deoxynucleoside monophosphates than does wild-type Pol ε, suggesting altered coordination between the polymerase and exonuclease active sites. The pol2-Y831A mutation suppresses the mutator effect of the pol2-4 mutation in the exonuclease active site that abolishes proofreading by Pol ε, as measured in haploid strain with the pol2-Y831A,4 double mutation. Analysis of mutation rates in diploid strains reveals that the pol2-Y831A allele is recessive to pol2-4. In addition, the mutation rates of strains with the pol2-4 mutation in combination with active site mutator mutations in Pol δ and Pol α suggest that Pol ε may proofread certain errors made by Pol α and Pol δ during replication in vivo. CONCLUSIONS: Our data suggest that Y831A replacement in Pol ε reduces replication fidelity and its participation in chromosomal replication, but without eliminating an additional function that is essential for viability. This suggests that other polymerases can substitute for certain functions of polymerase ε.
format Text
id pubmed-434536
institution National Center for Biotechnology Information
language English
publishDate 2004
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-4345362004-06-25 Evidence for interplay among yeast replicative DNA polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations Pavlov, Youri I Maki, Satoko Maki, Hisaji Kunkel, Thomas A BMC Biol Research Article BACKGROUND: DNA polymerase ε (Pol ε) is essential for S-phase replication, DNA damage repair and checkpoint control in yeast. A pol2-Y831A mutation leading to a tyrosine to alanine change in the Pol ε active site does not cause growth defects and confers a mutator phenotype that is normally subtle but strong in a mismatch repair-deficient strain. Here we investigate the mechanism responsible for the mutator effect. RESULTS: Purified four-subunit Y831A Pol ε turns over more deoxynucleoside triphosphates to deoxynucleoside monophosphates than does wild-type Pol ε, suggesting altered coordination between the polymerase and exonuclease active sites. The pol2-Y831A mutation suppresses the mutator effect of the pol2-4 mutation in the exonuclease active site that abolishes proofreading by Pol ε, as measured in haploid strain with the pol2-Y831A,4 double mutation. Analysis of mutation rates in diploid strains reveals that the pol2-Y831A allele is recessive to pol2-4. In addition, the mutation rates of strains with the pol2-4 mutation in combination with active site mutator mutations in Pol δ and Pol α suggest that Pol ε may proofread certain errors made by Pol α and Pol δ during replication in vivo. CONCLUSIONS: Our data suggest that Y831A replacement in Pol ε reduces replication fidelity and its participation in chromosomal replication, but without eliminating an additional function that is essential for viability. This suggests that other polymerases can substitute for certain functions of polymerase ε. BioMed Central 2004-05-26 /pmc/articles/PMC434536/ /pubmed/15163346 http://dx.doi.org/10.1186/1741-7007-2-11 Text en Copyright © 2004 Pavlov et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Research Article
Pavlov, Youri I
Maki, Satoko
Maki, Hisaji
Kunkel, Thomas A
Evidence for interplay among yeast replicative DNA polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations
title Evidence for interplay among yeast replicative DNA polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations
title_full Evidence for interplay among yeast replicative DNA polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations
title_fullStr Evidence for interplay among yeast replicative DNA polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations
title_full_unstemmed Evidence for interplay among yeast replicative DNA polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations
title_short Evidence for interplay among yeast replicative DNA polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations
title_sort evidence for interplay among yeast replicative dna polymerases alpha, delta and epsilon from studies of exonuclease and polymerase active site mutations
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC434536/
https://www.ncbi.nlm.nih.gov/pubmed/15163346
http://dx.doi.org/10.1186/1741-7007-2-11
work_keys_str_mv AT pavlovyourii evidenceforinterplayamongyeastreplicativednapolymerasesalphadeltaandepsilonfromstudiesofexonucleaseandpolymeraseactivesitemutations
AT makisatoko evidenceforinterplayamongyeastreplicativednapolymerasesalphadeltaandepsilonfromstudiesofexonucleaseandpolymeraseactivesitemutations
AT makihisaji evidenceforinterplayamongyeastreplicativednapolymerasesalphadeltaandepsilonfromstudiesofexonucleaseandpolymeraseactivesitemutations
AT kunkelthomasa evidenceforinterplayamongyeastreplicativednapolymerasesalphadeltaandepsilonfromstudiesofexonucleaseandpolymeraseactivesitemutations