Herpes simplex virus type 1 (HSV-1) uracil-DNA glycosylase: functional expression in Escherichia coli, biochemical characterization, and selective inhibition by 6-(p-n-octylanilino)uracil.
The Herpes simplex virus type 1 (HSV-1) uracil-DNA glycosylase (UDG) is encoded by the UL2 gene. The translation from the first putative start codon of UL2 predicts a polypeptide of 334 residues, while the translation from the second start codon predicts a polypeptide of 244 residues. We have cloned and expressed the two forms of UDG, by means of the prokaryotic expression vector pMAL-c2, and both of them were enzymatically active. Furthermore, the enzymatic properties of the recombinant UDGs and of the enzyme purified from HSV-1-infected cells were similar. The two UDG polypeptides have molecular weights of 27 and 37 kDa, respectively. The 37-kDa form of recombinant UDG is consistent with the reported molecular mass of 37 kDa for the enzyme purified from HSV-1-infected cells. Both recombinant UDGs were as sensitive as UDG purified from HSV-1-infected cells to 6-(p-n-octylanilino)uracil, the most potent of a series of uracil analogs that inhibit the viral enzyme.
Argnani
R
R
Institute of Microbiology, University of Ferrara, Italy.
Focher
F
F
Zucchini
S
S
Verri
A
A
Wright
G E
GE
Spadari
S
S
Manservigi
R
R
D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D045647:DNA Glycosylases; D017931:DNA Primers; D004260:DNA Repair; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D005814:Genes, Viral; D018259:Herpesvirus 1, Human / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009699:N-Glycosyl Hydrolases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D014498:Uracil / Q000031:analogs & derivatives* / Q000494:pharmacology; D051981:Uracil-DNA Glycosidase
C095137:6-(N-octylanilino)uracil; D003062:Codon; D017931:DNA Primers; D011994:Recombinant Proteins; D014498:Uracil; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D051981:Uracil-DNA Glycosidase