Mechanisms of base selection by human single-stranded selective monofunctional uracil-DNA glycosylase

Agus Darwanto, Jacob A. Theruvathu, James L. Sowers, Daniel K. Rogstad, Tod Pascal, William Goddard, Lawrence Sowers

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

hS,MUG1 (human single-stranded selective monofunctional uracil-DNA glyscosylase) is one of three glycosylases encoded within a small region of human chromosome 12. Those three glycosylases, UNG (uracil-DNA glycosylase), TDG (thymine-DNA glyscosylase), and hSMUG1, have in common the capacity to remove uracil from DNA. However, these glycosylases also repair other lesions and have distinct substrate preferences, indicating that they have potentially redundant but not overlapping physiological roles. The mechanisms by which these glycosylases locate and selectively remove target lesions are not well understood. In addition to uracil, hSMUG1 has been shown to remove some oxidized pyrimidines, suggesting a role in the repair of DNA oxidation damage. In this paper, we describe experiments in which a series of oligonucleotides containing purine and pyrimidine analogs have been used to probe mechanisms by which hSMUG1 distinguishes potential substrates. Our results indicate that the preference of hSMUG1 for mispaired uracil over uracil paired with adenine is best explained by the reduced stability of a duplex containing a mispair, consistent with previous reports with Escherichia coli mispaired uracil-DNA glycosylase. We have also extended the substrate range of hSMUG1 to include 5-carboxyuracil, the last in the series of damage products from thymine methyl group oxidation. The properties used by hSMUG1 to select damaged pyrimidines include the size and free energy of solvation of the 5-substituent but not electronic inductive properties. The observed distinct mechanisms of base selection demonstrated for members of the uracil glycosylase family help explain how considerable diversity in chemical lesion repair can be achieved.

Original languageEnglish (US)
Pages (from-to)15835-15846
Number of pages12
JournalJournal of Biological Chemistry
Volume284
Issue number23
DOIs
StatePublished - Jun 5 2009
Externally publishedYes

Fingerprint

Uracil-DNA Glycosidase
Uracil
Pyrimidines
Repair
Thymine
DNA
Substrates
Chromosomes, Human, Pair 12
Oxidation
Solvation
Human Chromosomes
Adenine
Chromosomes
Oligonucleotides
Escherichia coli
Free energy
DNA Damage

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Mechanisms of base selection by human single-stranded selective monofunctional uracil-DNA glycosylase. / Darwanto, Agus; Theruvathu, Jacob A.; Sowers, James L.; Rogstad, Daniel K.; Pascal, Tod; Goddard, William; Sowers, Lawrence.

In: Journal of Biological Chemistry, Vol. 284, No. 23, 05.06.2009, p. 15835-15846.

Research output: Contribution to journalArticle

Darwanto, A, Theruvathu, JA, Sowers, JL, Rogstad, DK, Pascal, T, Goddard, W & Sowers, L 2009, 'Mechanisms of base selection by human single-stranded selective monofunctional uracil-DNA glycosylase', Journal of Biological Chemistry, vol. 284, no. 23, pp. 15835-15846. https://doi.org/10.1074/jbc.M807846200
Darwanto, Agus ; Theruvathu, Jacob A. ; Sowers, James L. ; Rogstad, Daniel K. ; Pascal, Tod ; Goddard, William ; Sowers, Lawrence. / Mechanisms of base selection by human single-stranded selective monofunctional uracil-DNA glycosylase. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 23. pp. 15835-15846.
@article{e0db9280042b48fd9da60e83d369630a,
title = "Mechanisms of base selection by human single-stranded selective monofunctional uracil-DNA glycosylase",
abstract = "hS,MUG1 (human single-stranded selective monofunctional uracil-DNA glyscosylase) is one of three glycosylases encoded within a small region of human chromosome 12. Those three glycosylases, UNG (uracil-DNA glycosylase), TDG (thymine-DNA glyscosylase), and hSMUG1, have in common the capacity to remove uracil from DNA. However, these glycosylases also repair other lesions and have distinct substrate preferences, indicating that they have potentially redundant but not overlapping physiological roles. The mechanisms by which these glycosylases locate and selectively remove target lesions are not well understood. In addition to uracil, hSMUG1 has been shown to remove some oxidized pyrimidines, suggesting a role in the repair of DNA oxidation damage. In this paper, we describe experiments in which a series of oligonucleotides containing purine and pyrimidine analogs have been used to probe mechanisms by which hSMUG1 distinguishes potential substrates. Our results indicate that the preference of hSMUG1 for mispaired uracil over uracil paired with adenine is best explained by the reduced stability of a duplex containing a mispair, consistent with previous reports with Escherichia coli mispaired uracil-DNA glycosylase. We have also extended the substrate range of hSMUG1 to include 5-carboxyuracil, the last in the series of damage products from thymine methyl group oxidation. The properties used by hSMUG1 to select damaged pyrimidines include the size and free energy of solvation of the 5-substituent but not electronic inductive properties. The observed distinct mechanisms of base selection demonstrated for members of the uracil glycosylase family help explain how considerable diversity in chemical lesion repair can be achieved.",
author = "Agus Darwanto and Theruvathu, {Jacob A.} and Sowers, {James L.} and Rogstad, {Daniel K.} and Tod Pascal and William Goddard and Lawrence Sowers",
year = "2009",
month = "6",
day = "5",
doi = "10.1074/jbc.M807846200",
language = "English (US)",
volume = "284",
pages = "15835--15846",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "23",

}

TY - JOUR

T1 - Mechanisms of base selection by human single-stranded selective monofunctional uracil-DNA glycosylase

AU - Darwanto, Agus

AU - Theruvathu, Jacob A.

AU - Sowers, James L.

AU - Rogstad, Daniel K.

AU - Pascal, Tod

AU - Goddard, William

AU - Sowers, Lawrence

PY - 2009/6/5

Y1 - 2009/6/5

N2 - hS,MUG1 (human single-stranded selective monofunctional uracil-DNA glyscosylase) is one of three glycosylases encoded within a small region of human chromosome 12. Those three glycosylases, UNG (uracil-DNA glycosylase), TDG (thymine-DNA glyscosylase), and hSMUG1, have in common the capacity to remove uracil from DNA. However, these glycosylases also repair other lesions and have distinct substrate preferences, indicating that they have potentially redundant but not overlapping physiological roles. The mechanisms by which these glycosylases locate and selectively remove target lesions are not well understood. In addition to uracil, hSMUG1 has been shown to remove some oxidized pyrimidines, suggesting a role in the repair of DNA oxidation damage. In this paper, we describe experiments in which a series of oligonucleotides containing purine and pyrimidine analogs have been used to probe mechanisms by which hSMUG1 distinguishes potential substrates. Our results indicate that the preference of hSMUG1 for mispaired uracil over uracil paired with adenine is best explained by the reduced stability of a duplex containing a mispair, consistent with previous reports with Escherichia coli mispaired uracil-DNA glycosylase. We have also extended the substrate range of hSMUG1 to include 5-carboxyuracil, the last in the series of damage products from thymine methyl group oxidation. The properties used by hSMUG1 to select damaged pyrimidines include the size and free energy of solvation of the 5-substituent but not electronic inductive properties. The observed distinct mechanisms of base selection demonstrated for members of the uracil glycosylase family help explain how considerable diversity in chemical lesion repair can be achieved.

AB - hS,MUG1 (human single-stranded selective monofunctional uracil-DNA glyscosylase) is one of three glycosylases encoded within a small region of human chromosome 12. Those three glycosylases, UNG (uracil-DNA glycosylase), TDG (thymine-DNA glyscosylase), and hSMUG1, have in common the capacity to remove uracil from DNA. However, these glycosylases also repair other lesions and have distinct substrate preferences, indicating that they have potentially redundant but not overlapping physiological roles. The mechanisms by which these glycosylases locate and selectively remove target lesions are not well understood. In addition to uracil, hSMUG1 has been shown to remove some oxidized pyrimidines, suggesting a role in the repair of DNA oxidation damage. In this paper, we describe experiments in which a series of oligonucleotides containing purine and pyrimidine analogs have been used to probe mechanisms by which hSMUG1 distinguishes potential substrates. Our results indicate that the preference of hSMUG1 for mispaired uracil over uracil paired with adenine is best explained by the reduced stability of a duplex containing a mispair, consistent with previous reports with Escherichia coli mispaired uracil-DNA glycosylase. We have also extended the substrate range of hSMUG1 to include 5-carboxyuracil, the last in the series of damage products from thymine methyl group oxidation. The properties used by hSMUG1 to select damaged pyrimidines include the size and free energy of solvation of the 5-substituent but not electronic inductive properties. The observed distinct mechanisms of base selection demonstrated for members of the uracil glycosylase family help explain how considerable diversity in chemical lesion repair can be achieved.

UR - http://www.scopus.com/inward/record.url?scp=67650133665&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67650133665&partnerID=8YFLogxK

U2 - 10.1074/jbc.M807846200

DO - 10.1074/jbc.M807846200

M3 - Article

VL - 284

SP - 15835

EP - 15846

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 23

ER -