Crystal structure and mutational analysis of human uracil-DNA glycosylase: Structural basis for specificity and catalysis

Clifford D. Mol; Andrew S. Arvai; Geir Slupphaug; Bodil Kavli; Ingrun Alseth; Hans E. Krokan; John A. Tainer

doi:10.1016/0092-8674(95)90290-2

Crystal structure and mutational analysis of human uracil-DNA glycosylase: Structural basis for specificity and catalysis

Clifford D. Mol, Andrew S. Arvai, Geir Slupphaug, Bodil Kavli, Ingrun Alseth, Hans E. Krokan, John A. Tainer

Research output: Contribution to journal › Article › peer-review

347 Scopus citations

Abstract

Crystal structures of the DNA repair enzyme human uracil-DNA glycosylase (UDG), combined with mutational analysis, reveal the structural basis for the specificity of the enzyme. Within the classic α/β fold of UDG, sequence-conserved residues form a positively charged, active-site groove the width of duplex DNA, at the C-terminal edge of the central four-stranded parallel β sheet. In the UDG-6-aminouracil complex, uracil binds at the base of the groove within a rigid preformed pocket that confers selectivity for uracil over other bases by shape complementarity and by main chain and Asn-204 side chain hydrogen bonds. Main chain nitrogen atoms are positioned to stabilize the oxyanion intermediate generated by His-268 acting via nucleophilic attack or general base mechanisms. Specific binding of uracil flipped out from a DNA duplex provides a structural mechanism for damaged base recognition.

Original language	English (US)
Pages (from-to)	869-878
Number of pages	10
Journal	Cell
Volume	80
Issue number	6
DOIs	https://doi.org/10.1016/0092-8674(95)90290-2
State	Published - Mar 24 1995
Externally published	Yes

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

Access to Document

10.1016/0092-8674(95)90290-2

Cite this

@article{2297d60aaa144297bcdd8da532f9f10a,

title = "Crystal structure and mutational analysis of human uracil-DNA glycosylase: Structural basis for specificity and catalysis",

abstract = "Crystal structures of the DNA repair enzyme human uracil-DNA glycosylase (UDG), combined with mutational analysis, reveal the structural basis for the specificity of the enzyme. Within the classic α/β fold of UDG, sequence-conserved residues form a positively charged, active-site groove the width of duplex DNA, at the C-terminal edge of the central four-stranded parallel β sheet. In the UDG-6-aminouracil complex, uracil binds at the base of the groove within a rigid preformed pocket that confers selectivity for uracil over other bases by shape complementarity and by main chain and Asn-204 side chain hydrogen bonds. Main chain nitrogen atoms are positioned to stabilize the oxyanion intermediate generated by His-268 acting via nucleophilic attack or general base mechanisms. Specific binding of uracil flipped out from a DNA duplex provides a structural mechanism for damaged base recognition.",

author = "Mol, {Clifford D.} and Arvai, {Andrew S.} and Geir Slupphaug and Bodil Kavli and Ingrun Alseth and Krokan, {Hans E.} and Tainer, {John A.}",

note = "Funding Information: Correspondence should be addressed to J. A. T. The authors wish to thank E. D. Getzoff, T. J. Macke, H. E. Parge, R. P. Cunningham, G. E. O. Borgstahl, C. L. Fisher, K. J. Balch, and B. R. Crane for helpful advice and discussions and S. B. Petersen for discussions related to mutagenesis. This work was supported by grants from the Norwegian Cancer Society, the Norwegian Research Council, and the National Institutes of Health grant G M 46312.",

year = "1995",

month = mar,

day = "24",

doi = "10.1016/0092-8674(95)90290-2",

language = "English (US)",

volume = "80",

pages = "869--878",

journal = "Cell",

issn = "0092-8674",

publisher = "Cell Press",

number = "6",

}

TY - JOUR

T1 - Crystal structure and mutational analysis of human uracil-DNA glycosylase

T2 - Structural basis for specificity and catalysis

AU - Mol, Clifford D.

AU - Arvai, Andrew S.

AU - Slupphaug, Geir

AU - Kavli, Bodil

AU - Alseth, Ingrun

AU - Krokan, Hans E.

AU - Tainer, John A.

N1 - Funding Information: Correspondence should be addressed to J. A. T. The authors wish to thank E. D. Getzoff, T. J. Macke, H. E. Parge, R. P. Cunningham, G. E. O. Borgstahl, C. L. Fisher, K. J. Balch, and B. R. Crane for helpful advice and discussions and S. B. Petersen for discussions related to mutagenesis. This work was supported by grants from the Norwegian Cancer Society, the Norwegian Research Council, and the National Institutes of Health grant G M 46312.

PY - 1995/3/24

Y1 - 1995/3/24

N2 - Crystal structures of the DNA repair enzyme human uracil-DNA glycosylase (UDG), combined with mutational analysis, reveal the structural basis for the specificity of the enzyme. Within the classic α/β fold of UDG, sequence-conserved residues form a positively charged, active-site groove the width of duplex DNA, at the C-terminal edge of the central four-stranded parallel β sheet. In the UDG-6-aminouracil complex, uracil binds at the base of the groove within a rigid preformed pocket that confers selectivity for uracil over other bases by shape complementarity and by main chain and Asn-204 side chain hydrogen bonds. Main chain nitrogen atoms are positioned to stabilize the oxyanion intermediate generated by His-268 acting via nucleophilic attack or general base mechanisms. Specific binding of uracil flipped out from a DNA duplex provides a structural mechanism for damaged base recognition.

AB - Crystal structures of the DNA repair enzyme human uracil-DNA glycosylase (UDG), combined with mutational analysis, reveal the structural basis for the specificity of the enzyme. Within the classic α/β fold of UDG, sequence-conserved residues form a positively charged, active-site groove the width of duplex DNA, at the C-terminal edge of the central four-stranded parallel β sheet. In the UDG-6-aminouracil complex, uracil binds at the base of the groove within a rigid preformed pocket that confers selectivity for uracil over other bases by shape complementarity and by main chain and Asn-204 side chain hydrogen bonds. Main chain nitrogen atoms are positioned to stabilize the oxyanion intermediate generated by His-268 acting via nucleophilic attack or general base mechanisms. Specific binding of uracil flipped out from a DNA duplex provides a structural mechanism for damaged base recognition.

UR - http://www.scopus.com/inward/record.url?scp=0028934537&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028934537&partnerID=8YFLogxK

U2 - 10.1016/0092-8674(95)90290-2

DO - 10.1016/0092-8674(95)90290-2

M3 - Article

C2 - 7697717

AN - SCOPUS:0028934537

SN - 0092-8674

VL - 80

SP - 869

EP - 878

JO - Cell

JF - Cell

IS - 6

ER -

Crystal structure and mutational analysis of human uracil-DNA glycosylase: Structural basis for specificity and catalysis

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this