Enzymatic processing of uracil glycol, a major oxidative product of DNA cytosine

Andrei A. Purmal, Gary W. Lampman, Jeffrey P. Bond, Zafer Hatahet, Susan S. Wallace

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

A major stable oxidation product of DNA cytosine is uracil glycol (Ug). Because of the potential of Ug to be a strong premutagenic lesion, it is important to assess whether it is a blocking lesion to DNA polymerase as is its structural counterpart, thymine glycol (Tg), and to evaluate its pairing properties. Here, a series of oligonucleotides containing Ug or Tg were prepared and used as templates for a model enzyme, Escherichia coli DNA polymerase I Klenow fragment (exo-). During translesion DNA synthesis, Ug was bypassed more efficiently than Tg in all sequence contexts examined. Furthermore, only dAMP was incorporated opposite template Ug and Tg and the kinetic parameters of incorporation showed that dAMP was inserted opposite Ug more efficiently than opposite Tg. Ug opposite G and A was also recognized and removed in vitro by the E. coli DNA repair glycosylases, endonuclease III (endo III), endonuclease VIII (endo VIII), and formamidopyrimidine DNA glycosylase. The steady state kinetic parameters indicated that Ug was a better substrate for endo III and formamidopyrimidine DNA glycosylase than Tg; for endonuclease VIII, however, Tg was a better substrate.

Original languageEnglish (US)
Pages (from-to)10026-10035
Number of pages10
JournalJournal of Biological Chemistry
Volume273
Issue number16
DOIs
StatePublished - Apr 17 1998

Fingerprint

Cytosine
DNA
Processing
Deoxyribonuclease (Pyrimidine Dimer)
DNA-Formamidopyrimidine Glycosylase
DNA Polymerase I
Endonucleases
Kinetic parameters
Escherichia coli
DNA Glycosylases
uracil glycol
Substrates
DNA-Directed DNA Polymerase
thymine glycol
Oligonucleotides
Repair
Oxidation
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Purmal, Andrei A. ; Lampman, Gary W. ; Bond, Jeffrey P. ; Hatahet, Zafer ; Wallace, Susan S. / Enzymatic processing of uracil glycol, a major oxidative product of DNA cytosine. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 16. pp. 10026-10035.
@article{8dba4ffa9c424456832ae037d17b99f7,
title = "Enzymatic processing of uracil glycol, a major oxidative product of DNA cytosine",
abstract = "A major stable oxidation product of DNA cytosine is uracil glycol (Ug). Because of the potential of Ug to be a strong premutagenic lesion, it is important to assess whether it is a blocking lesion to DNA polymerase as is its structural counterpart, thymine glycol (Tg), and to evaluate its pairing properties. Here, a series of oligonucleotides containing Ug or Tg were prepared and used as templates for a model enzyme, Escherichia coli DNA polymerase I Klenow fragment (exo-). During translesion DNA synthesis, Ug was bypassed more efficiently than Tg in all sequence contexts examined. Furthermore, only dAMP was incorporated opposite template Ug and Tg and the kinetic parameters of incorporation showed that dAMP was inserted opposite Ug more efficiently than opposite Tg. Ug opposite G and A was also recognized and removed in vitro by the E. coli DNA repair glycosylases, endonuclease III (endo III), endonuclease VIII (endo VIII), and formamidopyrimidine DNA glycosylase. The steady state kinetic parameters indicated that Ug was a better substrate for endo III and formamidopyrimidine DNA glycosylase than Tg; for endonuclease VIII, however, Tg was a better substrate.",
author = "Purmal, {Andrei A.} and Lampman, {Gary W.} and Bond, {Jeffrey P.} and Zafer Hatahet and Wallace, {Susan S.}",
year = "1998",
month = "4",
day = "17",
doi = "10.1074/jbc.273.16.10026",
language = "English (US)",
volume = "273",
pages = "10026--10035",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "16",

}

Enzymatic processing of uracil glycol, a major oxidative product of DNA cytosine. / Purmal, Andrei A.; Lampman, Gary W.; Bond, Jeffrey P.; Hatahet, Zafer; Wallace, Susan S.

In: Journal of Biological Chemistry, Vol. 273, No. 16, 17.04.1998, p. 10026-10035.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Enzymatic processing of uracil glycol, a major oxidative product of DNA cytosine

AU - Purmal, Andrei A.

AU - Lampman, Gary W.

AU - Bond, Jeffrey P.

AU - Hatahet, Zafer

AU - Wallace, Susan S.

PY - 1998/4/17

Y1 - 1998/4/17

N2 - A major stable oxidation product of DNA cytosine is uracil glycol (Ug). Because of the potential of Ug to be a strong premutagenic lesion, it is important to assess whether it is a blocking lesion to DNA polymerase as is its structural counterpart, thymine glycol (Tg), and to evaluate its pairing properties. Here, a series of oligonucleotides containing Ug or Tg were prepared and used as templates for a model enzyme, Escherichia coli DNA polymerase I Klenow fragment (exo-). During translesion DNA synthesis, Ug was bypassed more efficiently than Tg in all sequence contexts examined. Furthermore, only dAMP was incorporated opposite template Ug and Tg and the kinetic parameters of incorporation showed that dAMP was inserted opposite Ug more efficiently than opposite Tg. Ug opposite G and A was also recognized and removed in vitro by the E. coli DNA repair glycosylases, endonuclease III (endo III), endonuclease VIII (endo VIII), and formamidopyrimidine DNA glycosylase. The steady state kinetic parameters indicated that Ug was a better substrate for endo III and formamidopyrimidine DNA glycosylase than Tg; for endonuclease VIII, however, Tg was a better substrate.

AB - A major stable oxidation product of DNA cytosine is uracil glycol (Ug). Because of the potential of Ug to be a strong premutagenic lesion, it is important to assess whether it is a blocking lesion to DNA polymerase as is its structural counterpart, thymine glycol (Tg), and to evaluate its pairing properties. Here, a series of oligonucleotides containing Ug or Tg were prepared and used as templates for a model enzyme, Escherichia coli DNA polymerase I Klenow fragment (exo-). During translesion DNA synthesis, Ug was bypassed more efficiently than Tg in all sequence contexts examined. Furthermore, only dAMP was incorporated opposite template Ug and Tg and the kinetic parameters of incorporation showed that dAMP was inserted opposite Ug more efficiently than opposite Tg. Ug opposite G and A was also recognized and removed in vitro by the E. coli DNA repair glycosylases, endonuclease III (endo III), endonuclease VIII (endo VIII), and formamidopyrimidine DNA glycosylase. The steady state kinetic parameters indicated that Ug was a better substrate for endo III and formamidopyrimidine DNA glycosylase than Tg; for endonuclease VIII, however, Tg was a better substrate.

UR - http://www.scopus.com/inward/record.url?scp=0032540240&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032540240&partnerID=8YFLogxK

U2 - 10.1074/jbc.273.16.10026

DO - 10.1074/jbc.273.16.10026

M3 - Article

VL - 273

SP - 10026

EP - 10035

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 16

ER -