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Environ Mol Mutagen. 2013 Oct;54(8):638-51. doi: 10.1002/em.21807. Epub 2013 Aug 1.

Mutation of the little finger domain in human DNA polymerase η alters fidelity when copying undamaged DNA.

Author information

1
Department of Environmental and Molecular Toxicology, North Carolina State University, Raleigh, North Carolina.

Abstract

DNA polymerase η (pol η) synthesizes past cyclobutane pyrimidine dimer and possibly 7,8-dihydro-8-oxoguanine (8-oxoG) lesions during DNA replication. Loss of pol η is associated with an increase in mutation rate, demonstrating its indispensable role in mutation suppression. It has been recently reported that β-strand 12 (amino acids 316-324) of the little finger region correctly positions the template strand with the catalytic core of the enzyme. The authors hypothesized that modification of β-strand 12 residues would disrupt correct enzyme-DNA alignment and alter pol η's activity and fidelity. To investigate this, the authors purified proteins containing the catalytic core of the polymerase, incorporated single amino acid changes to select β-strand 12 residues, and evaluated DNA synthesis activity for each pol η. Lesion bypass efficiencies and replication fidelities when copying DNA-containing cis-syn cyclobutane thymine-thymine dimer and 8-oxoG lesions were determined and compared with the corresponding values for the wild-type polymerase. The results confirm the importance of the β-strand in polymerase function and show that fidelity is most often altered when undamaged DNA is copied. Additionally, it is shown that DNA-protein contacts distal to the active site can significantly affect the fidelity of synthesis.

KEYWORDS:

DNA damage; lesion bypass; mutagenesis; replication fidelity

PMID:
23913529
PMCID:
PMC4120640
DOI:
10.1002/em.21807
[Indexed for MEDLINE]
Free PMC Article

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