Identification and characterization of duck enteritis virus dUTPase gene.
Deoxyuridine triphosphatase (dUTPase) is a ubiquitous and important enzyme that hydrolyzes dUTP to dUMP. Many viruses encode virus-specific dUTPase, which plays an essential role in maintaining the integrity of the viral DNA both by reducing the dUTP levels and by providing the substrate for the thymidylate synthase. A 1344-bp gene of duck enteritis virus (DEV) homologous to herpesviral dUTPase was first reported in this paper. The gene encodes a protein of 477 amino acids, with a predicted molecular mass of 49.7 kDa. Multiple sequence alignment suggested that DEV dUTPase was quite similar to other identified herpesviral dUTPase and functioned as a homotrimer. The five conserved motifs of DEV dUTPase with 3-1-2-4-5 arrangement have been recognized, and the phylogenetic analysis showed that DEV dUTPase was genetically close to the avian herpesvirus. Furthermore, RNA dot blot, western blot, and immunofluorescence analysis indicated that the enzyme was expressed at early and late stages after infection. Immunofluorescence also confirmed that DEV dUTPase localized in the cytoplasm of DEV-infected duck embryo fibroblasts as early as 4 hr postinfection (hpi). Later, the enzyme transferred from cytoplasm to nucleus at 8 hpi, and then reached its expression peak at 12 hpi, both in the cytoplasm and nucleus. The results suggested that the DEV dUTPase gene might be an early viral gene in DEV vitro infection and contribute to ensuring the fidelity of genome replication.
Zhao
Li-chan
LC
Avian Diseases Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Yaan, Sichuan, 625014, China.
Cheng
An-chun
AC
Wang
Ming-shu
MS
Yuan
Gui-ping
GP
Jia
Ren-Yong
RY
Zhou
Deng-chun
DC
Qi
Xue-Feng
XF
Ge
Han
H
Sun
Tao
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D004372:Ducks; D005814:Genes, Viral; D006564:Herpesviridae / Q000201:enzymology* / Q000235:genetics*; D006566:Herpesviridae Infections / Q000662:veterinary / Q000821:virology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011201:Poultry Diseases / Q000821:virology; D011755:Pyrophosphatases / Q000235:genetics* / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D012367:RNA, Viral; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase