Palm Mutants in DNA Polymerases α and η Alter DNA Replication Fidelity and Translesion Activity

Atsuko Niimi, Siripan Limsirichaikul, Shonen Yoshida, Shigenori Iwai, Chikahide Masutani, Fumio Hanaoka, Eric T. Kool, Yukihiro Nishiyama, Motoshi Suzuki

Research output: Contribution to journalArticle

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Abstract

We isolated active mutants in Saccharomyces cerevisiae DNA polymerase α that were associated with a defect in error discrimination. Among them, L868F DNA polymerase α has a spontaneous error frequency of 3 in 100 nucleotides and 570-fold lower replication fidelity than wild-type (WT) polymerase α. In vivo, mutant DNA polymerases confer a mutator phenotype and are synergistic with msh2 or msh6, suggesting that DNA polymerase α-dependent replication errors are recognized and repaired by mismatch repair. In vitro, L868F DNA polymerase α catalyzes efficient bypass of a cis-syn cyclobutane pyrimidine dimer, extending the 3′ T 26,000-fold more efficiently than the WT. Phe34 is equivalent to residue Leu868 in translesion DNA polymerase η, and the F34L mutant of S. cerevisiae DNA polymerase η has reduced translesion DNA synthesis activity in vitro. These data suggest that high-fidelity DNA synthesis by DNA polymerase α is required for genomic stability in yeast. The data also suggest that the phenylalanine and leucine residues in translesion and replicative DNA polymerases, respectively, might have played a role in the functional evolution of these enzyme classes.

Original languageEnglish
Pages (from-to)2734-2746
Number of pages13
JournalMolecular and Cellular Biology
Volume24
Issue number7
DOIs
Publication statusPublished - 01-04-2004
Externally publishedYes

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DNA-Directed DNA Polymerase
DNA Replication
Saccharomyces cerevisiae
Pyrimidine Dimers
DNA Mismatch Repair
Genomic Instability
DNA
Phenylalanine
Leucine
Nucleotides
Yeasts
Phenotype
Enzymes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Niimi, Atsuko ; Limsirichaikul, Siripan ; Yoshida, Shonen ; Iwai, Shigenori ; Masutani, Chikahide ; Hanaoka, Fumio ; Kool, Eric T. ; Nishiyama, Yukihiro ; Suzuki, Motoshi. / Palm Mutants in DNA Polymerases α and η Alter DNA Replication Fidelity and Translesion Activity. In: Molecular and Cellular Biology. 2004 ; Vol. 24, No. 7. pp. 2734-2746.
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Niimi, A, Limsirichaikul, S, Yoshida, S, Iwai, S, Masutani, C, Hanaoka, F, Kool, ET, Nishiyama, Y & Suzuki, M 2004, 'Palm Mutants in DNA Polymerases α and η Alter DNA Replication Fidelity and Translesion Activity', Molecular and Cellular Biology, vol. 24, no. 7, pp. 2734-2746. https://doi.org/10.1128/MCB.24.7.2734-2746.2004

Palm Mutants in DNA Polymerases α and η Alter DNA Replication Fidelity and Translesion Activity. / Niimi, Atsuko; Limsirichaikul, Siripan; Yoshida, Shonen; Iwai, Shigenori; Masutani, Chikahide; Hanaoka, Fumio; Kool, Eric T.; Nishiyama, Yukihiro; Suzuki, Motoshi.

In: Molecular and Cellular Biology, Vol. 24, No. 7, 01.04.2004, p. 2734-2746.

Research output: Contribution to journalArticle

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AU - Yoshida, Shonen

AU - Iwai, Shigenori

AU - Masutani, Chikahide

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AU - Suzuki, Motoshi

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