The UL2 open reading frame of bovine herpesvirus 1 encodes a uracil-DNA glycosylase.
Sequence analysis within the unique long segment of the bovine herpesvirus 1 (BHV-1) genome previously identified an open reading frame (ORF), designated UL2, whose deduced polypeptide of 204 amino acids contained a consensus uracil-DNA glycosylase (UDGase) signature sequence. To determine whether the BHV-1 UL2 ORF product has UDGase activity, we positioned the UL2 sequence down-stream of the T7 promoter on the vector pET-28b(+) and expressed it in Escherichia coli. Upon induction with isopropyl beta-D-thiogalactopyranoside these cells produced a 23-kDa protein, the molecular mass of which was in accordance with the prediction from the nucleotide sequence. A one-step purification procedure using nickel-chelating affinity chromatography resulted in a homogeneous preparation of this protein, which displayed specific UDGase activity in an in vitro enzyme assay. These results provide evidence that the BHV-1 UL2 gene does encode a UDGase.
Chung
Y T
YT
Department of Veterinary Medicine, National Chung-Hsing University, Taichung, Taiwan.
Hsu
W
W
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D016384:Consensus Sequence; D045647:DNA Glycosylases; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D007242:Herpesvirus 1, Bovine / Q000201:enzymology / Q000235:genetics*; D008970:Molecular Weight; D009699:N-Glycosyl Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016366:Open Reading Frames / Q000235:genetics; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D051981:Uracil-DNA Glycosidase
D003856:Deoxyuracil Nucleotides; D011994:Recombinant Proteins; C027078:deoxyuridine triphosphate; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D051981:Uracil-DNA Glycosidase