The Monomeric dUTPase from Epstein-Barr Virus Mimics Trimeric dUTPases

Abstract

The structure of Methanococcus jannaschii dUTPase was first determined by Johansson et al., 2003Johansson E. Bjornberg O. Nyman P.O. Larsen S. Structure of the bifunctional dCTP deaminase-dUTPase from Methanocaldococcus jannaschii and its relation to other homotrimeric dUTPases.J. Biol. Chem. 2003; 278: 27916-27922Crossref PubMed Scopus (20) Google Scholar. It is a bifunctional dCTP deaminase-dUTPase and not a pure dUTPase. We stated incorrectly in the introduction that β-herpesviruses, in particular human cytomegalovirus, code for dUTPases. In β-herpesviruses, the gene located at the position of the dUTPase does not code for a protein with dUTPase activity (Caposio et al., 2004Caposio P. Riera L. Hahn G. Landolfo S. Gribaudo G. Evidence that the Human Cytomegalovirus 46-kDa UL72 protein is not an active dUTPase but a late protein dispensable for replication in fibroblasts.Virology. 2004; 325: 264-276Crossref PubMed Scopus (26) Google Scholar), although it is structurally related to dUTPases (Davison and Stow, 2005Davison A.J. Stow N.D. New genes from old: redeployment of dUTPase by herpesviruses.J. Virol. 2005; 79: 12880-12892Crossref PubMed Scopus (63) Google Scholar, McGeehan et al., 2001McGeehan J.E. Depledge N.W. McGeoch D.J. Evolution of the dUTPase gene of mammalian and avian herpesviruses.Curr. Protein Pept. Sci. 2001; 2: 325-333Crossref PubMed Scopus (29) Google Scholar). Mutant HCMV virus deleted for this gene has a moderate growth defect (Dunn et al., 2003Dunn W. Chou C. Li H. Hai R. Patterson D. Stolc V. Zhu H. Liu F. Functional profiling of a human cytomegalovirus genome.Proc. Natl. Acad. Sci. USA. 2003; 100: 14223-14228Crossref PubMed Scopus (525) Google Scholar). We omitted a reference to the work of Sommer et al., 1996Sommer P. Kremmer E. Bier S. Konig S. Zalud P. Zeppezauer M. Jones J.F. Mueller-Lantzsch N. Grasser F.A. Cloning and expression of the Epstein-Barr virus-encoded dUTPase: patients with acute, reactivated or chronic virus infection develop antibodies against the enzyme.J. Gen. Virol. 1996; 77: 2795-2805Crossref PubMed Scopus (21) Google Scholar, who demonstrated the dUTPase activity of BLLF3 and determined a Km = 0.8 μM for EBV dUTPase. Previously, a dUTPase activity had been shown for EBV (Williams et al., 1985Williams M.V. Holliday J. Glaser R. Induction of a deoxyuridine triphosphate nucleotidohydrolase activity in Epstein-Barr virus-infected cells.Virology. 1985; 142: 326-333Crossref PubMed Scopus (25) Google Scholar) and the role of BLLF3 had been inferred from sequence homology with the HSV-1 UL50, for which dUTPase activity had been demonstrated (Williams, 1988Williams M.V. Herpes simplex virus-induced dUTPase: target site for antiviral chemotherapy.Virology. 1988; 166: 262-264Crossref PubMed Scopus (19) Google Scholar). The EBV dUTPase has been first detected in nasopharyngeal carcinoma by Nicholls et al., 1998Nicholls J.M. Sommer P. Kremmer E. Ong K.S. Fung K. Lee J.M. Ng M.H. Grasser F.A. A new lytic antibody, 7D6, detects Epstein-Barr virus dUTPase in nonkeratinizing undifferentiated nasopharyngeal carcinomas.Lab. Invest. 1998; 78: 1031-1032PubMed Google Scholar. The INDONESIA program used for Figure 1C has been written by D. Madsen, P. Johansson, and G. J. Kleywegt (unpublished work). The Monomeric dUTPase from Epstein-Barr Virus Mimics Trimeric dUTPasesTarbouriech et al.StructureSeptember, 2005In BriefDeoxyuridine 5′-triphosphate pyrophosphatases (dUTPases) are ubiquitous enzymes cleaving dUTP into dUMP and pyrophosphate. They occur as monomeric, dimeric, or trimeric molecules. The trimeric and monomeric enzymes both contain the same five characteristic sequence motifs but in a different order, whereas the dimeric enzymes are not homologous. Monomeric dUTPases only occur in herpesviruses, such as Epstein-Barr virus (EBV). Here, we describe the crystal structures of EBV dUTPase in complex with the product dUMP and a substrate analog α,β-imino-dUTP. Full-Text PDF Open Archive