Comparison of nucleotide interactions in water, proteins, and vacuum

Model for DNA polymerase fidelity

J. Petruska, Lawrence Sowers, M. F. Goodman

Research output: Contribution to journalArticle

135 Citations (Scopus)

Abstract

We propose a model for DNA polymerase fidelity in which free energy differences, ΔΔG, between matched and mismatched nucleotides are magnified at the enzyme's active site. Both hydrogen bonding and stacking components of the interaction energy are amplified, with the most profound effect being on the magnitude of hydrogen-bonding interactions. Magnification in ΔΔG values follows from the exclusion of water around base pairs in the active site cleft of the enzyme. After showing that base-pair dissociation energies calculated from hydrogen-bonding and base-stacking interactions in vacuo are greatly reduced by water, it is proposed that water removal results in a proportional restoration of these contributions to base pairing. Assuming ≃40% exclusion of surrounding water, one predicts magnified values of ΔΔG sufficient to account for polymerase insertion and proofreading fidelity, thereby avoiding the need to postulate additional active site constraints in order to select or reject nucleotides.

Original languageEnglish (US)
Pages (from-to)1559-1562
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume83
Issue number6
StatePublished - 1986
Externally publishedYes

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DNA-Directed DNA Polymerase
Vacuum
Hydrogen Bonding
Nucleotides
Base Pairing
Catalytic Domain
Water
Proteins
Enzymes

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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abstract = "We propose a model for DNA polymerase fidelity in which free energy differences, ΔΔG, between matched and mismatched nucleotides are magnified at the enzyme's active site. Both hydrogen bonding and stacking components of the interaction energy are amplified, with the most profound effect being on the magnitude of hydrogen-bonding interactions. Magnification in ΔΔG values follows from the exclusion of water around base pairs in the active site cleft of the enzyme. After showing that base-pair dissociation energies calculated from hydrogen-bonding and base-stacking interactions in vacuo are greatly reduced by water, it is proposed that water removal results in a proportional restoration of these contributions to base pairing. Assuming ≃40{\%} exclusion of surrounding water, one predicts magnified values of ΔΔG sufficient to account for polymerase insertion and proofreading fidelity, thereby avoiding the need to postulate additional active site constraints in order to select or reject nucleotides.",
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AB - We propose a model for DNA polymerase fidelity in which free energy differences, ΔΔG, between matched and mismatched nucleotides are magnified at the enzyme's active site. Both hydrogen bonding and stacking components of the interaction energy are amplified, with the most profound effect being on the magnitude of hydrogen-bonding interactions. Magnification in ΔΔG values follows from the exclusion of water around base pairs in the active site cleft of the enzyme. After showing that base-pair dissociation energies calculated from hydrogen-bonding and base-stacking interactions in vacuo are greatly reduced by water, it is proposed that water removal results in a proportional restoration of these contributions to base pairing. Assuming ≃40% exclusion of surrounding water, one predicts magnified values of ΔΔG sufficient to account for polymerase insertion and proofreading fidelity, thereby avoiding the need to postulate additional active site constraints in order to select or reject nucleotides.

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