The Mechanism of Nucleotide Incorporation by Human DNA Polymerase η Differs from That of the Yeast Enzyme

Washington, M. Todd; Johnson, Robert E.; Prakash, Louise; Prakash, Satya

doi:10.1128/mcb.23.22.8316-8322.2003

Cited by 45DNA Repair MechanismsDNA and Nucleic Acid ChemistryEnzyme Structure and Function

The Mechanism of Nucleotide Incorporation by Human DNA Polymerase η Differs from That of the Yeast Enzyme

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Washington, M. Todd, Johnson, Robert E., Prakash, Louise, Prakash, Satya·2003·Molecular and Cellular Biology

DOI 10.1128/mcb.23.22.8316-8322.2003PMC PMC262418

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Abstract

DNA polymerase eta (Poleta) catalyzes the efficient and accurate synthesis of DNA opposite cyclobutane pyrimidine dimers, and inactivation of Poleta in humans causes the cancer-prone syndrome, the variant form of xeroderma pigmentosum. Pre-steady-state kinetic studies of yeast Poleta have indicated that the low level of fidelity of this enzyme results from a poorly discriminating induced-fit mechanism. Here we examine the mechanistic basis of the low level of fidelity of human Poleta. Because the human and yeast enzymes behave similarly under steady-state conditions, we expected these enzymes to utilize similar mechanisms of nucleotide incorporation. Surprisingly, however, we find that human Poleta differs from the yeast enzyme in several important respects. The human enzyme has a 50-fold-faster rate of nucleotide incorporation than the yeast enzyme but binds the nucleotide with an approximately 50-fold-lower level of affinity. This lower level of binding affinity might provide a means of regulation whereby the human enzyme remains relatively inactive except when the cellular deoxynucleoside triphosphate concentrations are high, as may occur during DNA damage, thereby avoiding the mutagenic consequences arising from the inadvertent action of this enzyme during normal DNA replication.

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These multiplet mutations are likely the consequence of error-prone bypass polymerization during translesion synthesis and are consistent with processivity of bypass polymerases persisting for several nucleotides after the bypass-requiring blocking lesion.
“These multiplet mutations are likely the consequence of error-prone bypass polymerization during translesion synthesis and are consistent with the processivity of bypass polymerases persisting for several nucleotides after the bypass-requiring blocking lesion (39 -41).”
From Why Cockayne syndrome patients do not get cancer despite their DNA repair deficiency (2016) · cites this paper as [40]

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Background
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BackgroundInfluential
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BackgroundMethodResult
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Method
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Background
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Background
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Authors: Washington, M. T., Johnson, R. E., Prakash, L., & Prakash, S.
Affiliations: The University of Texas Medical Branch at Galveston (US)
ORCID: Washington, M. Todd
Johnson, Robert E.
Prakash, Louise
Prakash, Satya
Venue: Molecular and Cellular Biology
Publisher: Informa UK Limited
Published: 2003-11-01
Volume: 23
Issue: 22
Pages: 8316-8322
ISSN: 0270-7306 (ISSN-L)
1098-5549
DOI: 10.1128/mcb.23.22.8316-8322.2003
PMC: PMC262418
OpenAlex: W2136817602
Cited by: 45
References: 0
Citations by year: 2025: 1 · 2024: 1 · 2021: 1 · 2019: 4 · 2018: 3 · 2016: 3 · 2015: 4 · 2014: 1 · 2013: 1 · 2012: 2
FWCI: 1.15
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Molecular and Cellular Biology (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health (5r01gm019261-30)
National Institute of General Medical Sciences (GM-19261)
National Institute of Environmental Health Sciences (R01ES012411, ES 012411)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
Keywords: DNA polymerase eta, xeroderma pigmentosum, cyclobutane pyrimidine dimers, DNA repair, mutagenesis, induced fit, nucleotide binding affinity, enzyme regulation, DNA replication fidelity
MeSH terms: Y-Family DNA Polymerases, Base Sequence, DNA Damage, DNA-Directed DNA Polymerase, Humans, Kinetics, Oligodeoxyribonucleotides, Saccharomyces cerevisiae, Species Specificity, Substrate Specificity, Xeroderma Pigmentosum, Molecular Structure, Sequence Deletion

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The Mechanism of Nucleotide Incorporation by Human DNA Polymerase η Differs from That of the Yeast Enzyme

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