Requirement of Watson-Crick hydrogen bonding for DNA synthesis by yeast DNA polymerase η

M. Todd Washington, Sandra A. Helquist, Eric T. Kool, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Classical high-fidelity DNA polymerases discriminate between the correct and incorrect nucleotides by using geometric constraints imposed by the tight fit of the active site with the incipient base pair. Consequently, Watson-Crick (W-C) hydrogen bonding between the bases is not required for the efficiency and accuracy of DNA synthesis by these polymerases. DNA polymerase η (Polη) is a low-fidelity enzyme able to replicate through DNA lesions. Using difluorotoluene, a nonpolar isosteric analog of thymine unable to form W-C hydrogen bonds with adenine, we found that the efficiency and accuracy of nucleotide incorporation by Polη are severely impaired. From these observations, we suggest that W-C hydrogen bonding is required for DNA synthesis by Polη; in this regard, Polη differs strikingly from classical high-fidelity DNA polymerases.

Original languageEnglish (US)
Pages (from-to)5107-5112
Number of pages6
JournalMolecular and Cellular Biology
Volume23
Issue number14
DOIs
StatePublished - Jul 2003

Fingerprint

DNA-Directed DNA Polymerase
Hydrogen Bonding
Yeasts
DNA
Nucleotides
Thymine
Adenine
Base Pairing
Hydrogen
Catalytic Domain
Enzymes

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Requirement of Watson-Crick hydrogen bonding for DNA synthesis by yeast DNA polymerase η. / Washington, M. Todd; Helquist, Sandra A.; Kool, Eric T.; Prakash, Louise; Prakash, Satya.

In: Molecular and Cellular Biology, Vol. 23, No. 14, 07.2003, p. 5107-5112.

Research output: Contribution to journalArticle

Washington, M. Todd ; Helquist, Sandra A. ; Kool, Eric T. ; Prakash, Louise ; Prakash, Satya. / Requirement of Watson-Crick hydrogen bonding for DNA synthesis by yeast DNA polymerase η. In: Molecular and Cellular Biology. 2003 ; Vol. 23, No. 14. pp. 5107-5112.
@article{2ab977a1f6e8489aa765987c7db0500b,
title = "Requirement of Watson-Crick hydrogen bonding for DNA synthesis by yeast DNA polymerase η",
abstract = "Classical high-fidelity DNA polymerases discriminate between the correct and incorrect nucleotides by using geometric constraints imposed by the tight fit of the active site with the incipient base pair. Consequently, Watson-Crick (W-C) hydrogen bonding between the bases is not required for the efficiency and accuracy of DNA synthesis by these polymerases. DNA polymerase η (Polη) is a low-fidelity enzyme able to replicate through DNA lesions. Using difluorotoluene, a nonpolar isosteric analog of thymine unable to form W-C hydrogen bonds with adenine, we found that the efficiency and accuracy of nucleotide incorporation by Polη are severely impaired. From these observations, we suggest that W-C hydrogen bonding is required for DNA synthesis by Polη; in this regard, Polη differs strikingly from classical high-fidelity DNA polymerases.",
author = "Washington, {M. Todd} and Helquist, {Sandra A.} and Kool, {Eric T.} and Louise Prakash and Satya Prakash",
year = "2003",
month = "7",
doi = "10.1128/MCB.23.14.5107-5112.2003",
language = "English (US)",
volume = "23",
pages = "5107--5112",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "14",

}

TY - JOUR

T1 - Requirement of Watson-Crick hydrogen bonding for DNA synthesis by yeast DNA polymerase η

AU - Washington, M. Todd

AU - Helquist, Sandra A.

AU - Kool, Eric T.

AU - Prakash, Louise

AU - Prakash, Satya

PY - 2003/7

Y1 - 2003/7

N2 - Classical high-fidelity DNA polymerases discriminate between the correct and incorrect nucleotides by using geometric constraints imposed by the tight fit of the active site with the incipient base pair. Consequently, Watson-Crick (W-C) hydrogen bonding between the bases is not required for the efficiency and accuracy of DNA synthesis by these polymerases. DNA polymerase η (Polη) is a low-fidelity enzyme able to replicate through DNA lesions. Using difluorotoluene, a nonpolar isosteric analog of thymine unable to form W-C hydrogen bonds with adenine, we found that the efficiency and accuracy of nucleotide incorporation by Polη are severely impaired. From these observations, we suggest that W-C hydrogen bonding is required for DNA synthesis by Polη; in this regard, Polη differs strikingly from classical high-fidelity DNA polymerases.

AB - Classical high-fidelity DNA polymerases discriminate between the correct and incorrect nucleotides by using geometric constraints imposed by the tight fit of the active site with the incipient base pair. Consequently, Watson-Crick (W-C) hydrogen bonding between the bases is not required for the efficiency and accuracy of DNA synthesis by these polymerases. DNA polymerase η (Polη) is a low-fidelity enzyme able to replicate through DNA lesions. Using difluorotoluene, a nonpolar isosteric analog of thymine unable to form W-C hydrogen bonds with adenine, we found that the efficiency and accuracy of nucleotide incorporation by Polη are severely impaired. From these observations, we suggest that W-C hydrogen bonding is required for DNA synthesis by Polη; in this regard, Polη differs strikingly from classical high-fidelity DNA polymerases.

UR - http://www.scopus.com/inward/record.url?scp=0037773539&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037773539&partnerID=8YFLogxK

U2 - 10.1128/MCB.23.14.5107-5112.2003

DO - 10.1128/MCB.23.14.5107-5112.2003

M3 - Article

VL - 23

SP - 5107

EP - 5112

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 14

ER -