A rapid reaction analysis of uracil DNA glycosylase indicates an active mechanism of base flipping
Author(s)
Bellamy, SRW
Krusong, K
Baldwin, GS
Type
Journal Article
Abstract
Uracil DNA glycosylase (UNG) is the primary enzyme
for the removal of uracil from the genome of many
organisms. A key question is how the enzyme is
able to scan large quantities of DNA in search of
aberrant uracil residues. Central to this is the
mechanism by which it flips the target nucleotide
out of the DNA helix and into the enzyme-active site.
Both active and passive mechanisms have been
proposed. Here, we report a rapid kinetic analysis
using two fluorescent chromophores to temporally
resolve DNA binding and base-flipping with DNA
substrates of different sequences. This study
demonstrates the importance of the protein–DNA
interface in the search process and indicates an
active mechanism by which UNG glycosylase
searches for uracil residues.
for the removal of uracil from the genome of many
organisms. A key question is how the enzyme is
able to scan large quantities of DNA in search of
aberrant uracil residues. Central to this is the
mechanism by which it flips the target nucleotide
out of the DNA helix and into the enzyme-active site.
Both active and passive mechanisms have been
proposed. Here, we report a rapid kinetic analysis
using two fluorescent chromophores to temporally
resolve DNA binding and base-flipping with DNA
substrates of different sequences. This study
demonstrates the importance of the protein–DNA
interface in the search process and indicates an
active mechanism by which UNG glycosylase
searches for uracil residues.
Date Issued
2007-01-01
Date Acceptance
2007-01-02
Citation
Nucleic Acids Research, 2007, 35 (5), pp.1478-1487
ISSN
1362-4962
Publisher
Oxford University Press (OUP)
Start Page
1478
End Page
1487
Journal / Book Title
Nucleic Acids Research
Volume
35
Issue
5
Copyright Statement
© 2007 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/
by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/
by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
BIOCHEMISTRY & MOLECULAR BIOLOGY
ESCHERICHIA-COLI
STRUCTURAL BASIS
EXCISION-REPAIR
CRYSTAL-STRUCTURE
METHYLTRANSFERASE
FLUORESCENCE
RECOGNITION
BINDING
2-AMINOPURINE
SPECIFICITY
Publication Status
Published