Argnani R, Focher F, Zucchini S, Verri A, Wright G E, Spadari S, Manservigi R
Institute of Microbiology, University of Ferrara, Italy.
Virology. 1995 Aug 1;211(1):307-11. doi: 10.1006/viro.1995.1406.
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.
单纯疱疹病毒1型（HSV-1）尿嘧啶-DNA糖基化酶（UDG）由UL2基因编码。从UL2的第一个推定起始密码子开始翻译，预测会产生一个由334个残基组成的多肽，而从第二个起始密码子开始翻译，则预测会产生一个由244个残基组成的多肽。我们通过原核表达载体pMAL-c2克隆并表达了两种形式的UDG，且它们都具有酶活性。此外，重组UDG和从感染HSV-1的细胞中纯化得到的酶的酶学性质相似。这两种UDG多肽的分子量分别为27 kDa和37 kDa。重组UDG的37 kDa形式与从感染HSV-1的细胞中纯化得到的该酶报道的分子量37 kDa一致。两种重组UDG对6-（对正辛基苯胺基）尿嘧啶的敏感性与从感染HSV-1的细胞中纯化得到的UDG相同，6-（对正辛基苯胺基）尿嘧啶是一系列抑制该病毒酶的尿嘧啶类似物中最有效的一种。
