Literature summary for 3.2.2.27 extracted from

Schormann, N.; Grigorian, A.; Samal, A; Krishnan, R.; DeLucas, L.; Chattopadhyay, D.
Crystal structure of vaccinia virus uracil-DNA glycosylase reveals dimeric assembly (2007), BMC Struct. Biol., 7, 45.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of His-tagged UDG in Escherichia coli strain BL21(DE3), sequence comparison	Vaccinia virus

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant thrombin-cleaved, detagged enzyme, crystallization in two different conditions, condition 1: 5% PEG 6000, 7.5% MPD, 0.1 M HEPES, pH 7.25, at 4°C; condition 2: 5% PEG 3000, 0.1 M NaCl, 0.1 M HEPES, pH 7.5, at 4°C, 1-3 days, or protein purified in the one-step procedure, crystallized by hanging-drop vapor diffusion, pH 7.3, 100 mM HEPES buffer, pH 7.25, 12% glycerol, and 1.5 M ammonium sulfate as precipitant, improvement of crystallization by microseeding, X-ray diffraction structure determination and analysis at 2.4 A resolution, heavy atom labeling and molecular replacement, modelling	Vaccinia virus

Crystallization (Comment)

Organism

purified recombinant thrombin-cleaved, detagged enzyme, crystallization in two different conditions, condition 1: 5% PEG 6000, 7.5% MPD, 0.1 M HEPES, pH 7.25, at 4°C; condition 2: 5% PEG 3000, 0.1 M NaCl, 0.1 M HEPES, pH 7.5, at 4°C, 1-3 days, or protein purified in the one-step procedure, crystallized by hanging-drop vapor diffusion, pH 7.3, 100 mM HEPES buffer, pH 7.25, 12% glycerol, and 1.5 M ammonium sulfate as precipitant, improvement of crystallization by microseeding, X-ray diffraction structure determination and analysis at 2.4 A resolution, heavy atom labeling and molecular replacement, modelling

Vaccinia virus

Protein Variants

Protein Variants	Comment	Organism
G179R	structure modelling of the temperature-sensitive mutant Dts30, overview	Vaccinia virus
L110F	structure modelling of the temperature-sensitive mutant Dts27, overview	Vaccinia virus

Inhibitors

Inhibitors	Comment	Organism	Structure
MgCl2	strong inhibition	Vaccinia virus
additional information	no inhibition of Vaccinia virus UDG by uracil-DNA glycosylase inhibitor protein, Ugi, from Bacillus subtilis bacteriophages PBS1 and PBS2	Vaccinia virus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0005	-	uracil-mismatched single-stranded DNA	-	Vaccinia virus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
57000	-	dynamic light scattering and gel filtration	Vaccinia virus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
uracil-mismatched double-stranded DNA + H2O	Vaccinia virus	UDG catalyzes excision of uracil from DNA. The viral UDG plays an essential role in viral replication as a component of the DNA polymerase processivity factor. It adopts a catalysis-independent role in DNA replication that involves interaction with a viral protein, A20, to form the processivity factor. UDG-A20 association is essential for assembling of the processive DNA polymerase complex, overview	uracil + double-stranded DNA with abasic site	-	?

Organism

Organism	UniProt	Comment	Textmining
Vaccinia virus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged UDG from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration, cleavage of the His-tag by thrombin	Vaccinia virus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
uracil-mismatched double-stranded DNA + H2O	UDG catalyzes excision of uracil from DNA. The viral UDG plays an essential role in viral replication as a component of the DNA polymerase processivity factor. It adopts a catalysis-independent role in DNA replication that involves interaction with a viral protein, A20, to form the processivity factor. UDG-A20 association is essential for assembling of the processive DNA polymerase complex, overview	Vaccinia virus	uracil + double-stranded DNA with abasic site	-	?
uracil-mismatched single-stranded DNA + H2O	The viral UDG enzyme is highly specific for uracil and preferentially excises uracil when present in single stranded DNA, the reaction mechanism of the viral enzyme is different from the human host enzyme, active site structure and motifs, modelling, overview	Vaccinia virus	uracil + single-stranded DNA with abasic site	-	?

Subunits

Subunits	Comment	Organism
dimer	dynamic light scattering and gel filtration	Vaccinia virus
More	the enzyme differs from UDGs of other species in its primary, secondary and tertiary structure, through modifications of otherwise conserved active site motifs, assembly and protein-protein interactions, overview	Vaccinia virus

Synonyms

Synonyms	Comment	Organism
UDG	-	Vaccinia virus