1887

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Volume 85, Issue 10

Identification and characterization of a novel gene of grouper iridovirus encoding a purine nucleoside phosphorylase Free

Abstract

Purine nucleoside phosphorylase (PNP) is a key enzyme in the purine salvage pathway. It catalyses the reversible phosphorolysis of purine (2′-deoxy)ribonucleosides to free bases and (2′-deoxy)ribose 1-phosphates. Here, a novel piscine viral gene that was identified from grouper iridovirus (GIV), a causative agent of an epizootic fish disease, is reported. This putative GIV gene encodes a protein of 285 aa with a predicted molecular mass of 30 332 Da and shows high similarity to the human gene. Northern and Western blot analyses of GIV-infected grouper kidney (GK) cells revealed that PNP expression increased in cells with time from 6 h post-infection. Immunocytochemistry localized GIV PNP in the cytoplasm of GIV-infected host cells. PNP–EGFP fusion protein was also observed in the cytoplasm of PNP–EGFP reporter construct-transfected GK and HeLa cells. From HPLC analysis, the recombinant GIV PNP protein was shown to catalyse the reversible phosphorolysis of purine nucleosides and could accept guanosine, inosine and adenosine as substrates. In conclusion, this is the first report of a viral PNP with enzymic activity.

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2004-10-01
2024-05-03
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References

  1. Altschul S. F., Madden T. L., Schäffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402 [CrossRef]
     [Google Scholar]
  2. Alvarez F., Ghérardi A., Nebois P., Sarciron M.-E., Pétavy A.-F., Walchshofer N. 2002; Benzimidazole-4,7-diones as inhibitors of protozoal ( Toxoplasma gondii ) purine nucleoside phosphorylase. Bioorg Med Chem Lett 12:977–979 [CrossRef]
     [Google Scholar]
  3. Bantia S., Miller P. J., Parker C. D. 7 other authors 2001; Purine nucleoside phosphorylase inhibitor BCX-1777 (Immucillin-H) – a novel potent and orally active immunosuppressive agent. Int Immunopharmacol 1:1199–1210 [CrossRef]
     [Google Scholar]
  4. Beaud G. 1995; Vaccinia virus DNA replication: a short review. Biochimie 77:774–779 [CrossRef]
     [Google Scholar]
  5. Binns M. M., Boursnell M. E. G., Skinner M. A. 1992; Gene translocations in poxviruses: the fowlpox virus thymidine kinase gene is flanked by 15 bp direct repeats and occupies the locus which in vaccinia virus is occupied by the ribonucleotide reductase large subunit gene. Virus Res 24:161–172 [CrossRef]
     [Google Scholar]
  6. Boehmer P. E., Lehman I. R. 1997; Herpes simplex virus DNA replication. Annu Rev Biochem 66:347–384 [CrossRef]
     [Google Scholar]
  7. Breidbach T., Krauth-Siegel R. L., Steverding D. 2000; Ribonucleotide reductase is regulated via the R2 subunit during the life cycle of Trypanosoma brucei . FEBS Lett 473:212–216 [CrossRef]
     [Google Scholar]
  8. Bzowska A., Luić M., Schröder W., Shugar D., Saenger W., Koellner G. 1995; Calf spleen purine nucleoside phosphorylase: purification, sequence and crystal structure of its complex with an N (7)-acycloguanosine inhibitor. FEBS Lett 367:214–218 [CrossRef]
     [Google Scholar]
  9. Bzowska A., Kulikowska E., Shugar D. 2000; Purine nucleoside phosphorylases: properties, functions, and clinical aspects. Pharmacol Ther 88:349–425 [CrossRef]
     [Google Scholar]
  10. Chen S.-H., Cook W. J., Grove K. L., Coen D. M. 1998; Human thymidine kinase can functionally replace herpes simplex virus type 1 thymidine kinase for viral replication in mouse sensory ganglia and reactivation from latency upon explant. J Virol 72:6710–6715
     [Google Scholar]
  11. Chen X. H., Lin K. B., Wang X. W. 2003; Outbreaks of an iridovirus disease in maricultured large yellow croaker, Larimichthys crocea (Richardson), in China. J Fish Dis 26:615–619 [CrossRef]
     [Google Scholar]
  12. Chou H.-Y., Hsu C.-C., Peng T.-Y. 1998; Isolation and characterization of a pathogenic iridovirus from cultured grouper ( Epinephelus sp.) in Taiwan. Fish Pathol 33:201–206 [CrossRef]
     [Google Scholar]
  13. Delius H., Darai G., Flügel R. M. 1984; DNA analysis of insect iridescent virus 6: evidence for circular permutation and terminal redundancy. J Virol 49:609–614
     [Google Scholar]
  14. Erion M. D., Takabayashi K., Smith H. B., Kessi J., Wagner S., Hönger S., Shames S. L., Ealick S. E. 1997; Purine nucleoside phosphorylase. 1. Structure-function studies. Biochemistry 36:11725–11734 [CrossRef]
     [Google Scholar]
  15. Gadi V. K., Alexander S. D., Kudlow J. E., Allan P., Parker W. B., Sorscher E. J. 2000; In vivo sensitization of ovarian tumors to chemotherapy by expression of E. coli purine nucleoside phosphorylase in a small fraction of cells. Gene Ther 7:1738–1743 [CrossRef]
     [Google Scholar]
  16. Gadi V. K., Alexander S. D., Waud W. R., Allan P. W., Parker W. B., Sorscher E. J. 2003; A long-acting suicide gene toxin, 6-methylpurine, inhibits slow growing tumors after a single administration. J Pharmacol Exp Ther 304:1280–1284
     [Google Scholar]
  17. Giblett E. R., Ammann A. J., Sandman R., Wara D. W., Diamond L. K. 1975; Nucleoside-phosphorylase deficiency in a child with severely defective T-cell immunity and normal B-cell immunity. Lancet 305:1010–1013 [CrossRef]
     [Google Scholar]
  18. Gregus Z., Németi B. 2002; Purine nucleoside phosphorylase as a cytosolic arsenate reductase. Toxicol Sci 70:13–19 [CrossRef]
     [Google Scholar]
  19. He J. G., Wang S. P., Zeng K., Huang Z. J., Chan S.-M. 2000; Systemic disease caused by an iridovirus-like agent in cultured mandarinfish, Siniperca chuatsi (Basilewsky), in China. J Fish Dis 23:219–222 [CrossRef]
     [Google Scholar]
  20. He J. G., Deng M., Weng S. P., Li Z., Zhou S. Y., Long Q. X., Wang X. Z., Chan S.-M. 2001; Complete genome analysis of the mandarin fish infectious spleen and kidney necrosis iridovirus. Virology 291:126–139 [CrossRef]
     [Google Scholar]
  21. He J. G., Lü L., Deng M. 7 other authors 2002; Sequence analysis of the complete genome of an iridovirus isolated from the tiger frog. Virology 292:185–197 [CrossRef]
     [Google Scholar]
  22. Hughes B. W., King S. A., Allan P. W., Parker W. B., Sorscher E. J. 1998; Cell to cell contact is not required for bystander cell killing by Escherichia coli purine nucleoside phosphorylase. J Biol Chem 273:2322–2328 [CrossRef]
     [Google Scholar]
  23. Hwang H.-Y., Ullman B. 1997; Genetic analysis of purine metabolism in Leishmania donovani . J Biol Chem 272:19488–19496 [CrossRef]
     [Google Scholar]
  24. Inouye K., Yamano K., Maeno Y., Nakajima K., Matsuoka M., Wada Y., Sorimachi M. 1992; Iridovirus infection of cultured red sea bream, Pagrus major . Fish Pathol 27:19–27 [CrossRef]
     [Google Scholar]
  25. Jakob N. J., Müller K., Bahr U., Darai G. 2001; Analysis of the first complete DNA sequence of an invertebrate iridovirus: coding strategy of the genome of Chilo iridescent virus. Virology 286:182–196 [CrossRef]
     [Google Scholar]
  26. Jancovich J. K., Mao J., Chinchar V. G. 8 other authors 2003; Genomic sequence of a ranavirus (family Iridoviridae ) associated with salamander mortalities in North America. Virology 316:90–103 [CrossRef]
     [Google Scholar]
  27. Jenuth J. P., Mangat R. K., Snyder F. F. 1993; cDNA sequence of four purine nucleoside phosphorylase (Np) alleles in the mouse. Mamm Genome 4:598–603 [CrossRef]
     [Google Scholar]
  28. Jung S. J., Oh M. J. 2000; Iridovirus-like infection associated with high mortalities of striped beakperch, Oplegnathus fasciatus (Temminck et Schlegel), in southern coastal areas of the Korean peninsula. J Fish Dis 23:223–226 [CrossRef]
     [Google Scholar]
  29. Kicska G. A., Tyler P. C., Evans G. B., Furneaux R. H., Kim K., Schramm V. L. 2002a; Transition state analogue inhibitors of purine nucleoside phosphorylase from Plasmodium falciparum . J Biol Chem 277:3219–3225 [CrossRef]
     [Google Scholar]
  30. Kicska G. A., Tyler P. C., Evans G. B., Furneaux R. H., Schramm V. L., Kim K. 2002b; Purine-less death in Plasmodium falciparum induced by immucillin-H, a transition state analogue of purine nucleoside phosphorylase. J Biol Chem 277:3226–3231 [CrossRef]
     [Google Scholar]
  31. Krug E. C., Marr J. J., Berens R. L. 1989; Purine metabolism in Toxoplasma gondii . J Biol Chem 264:10601–10607
     [Google Scholar]
  32. Lai Y.-S., Murali S., Ju H.-Y., Wu M.-F., Guo I.-C., Chen S.-C., Fang K., Chang C.-Y. 2000; Two iridovirus-susceptible cell lines established from kidney and liver of grouper, Epinephelus awoara Temminck & Schlegel. and partial characterization of grouper iridovirus. J Fish Dis 23379–388 [CrossRef]
     [Google Scholar]
  33. Lai Y.-S., John J. A. C., Lin C.-H., Guo I.-C., Chen S.-C., Fang K., Lin C.-H., Chang C.-Y. 2003; Establishment of cell lines from a tropical grouper, Epinephelus awoara (Temminck & Schlegel. ) and their susceptibility to grouper irido- and nodaviruses. J Fish Dis 2631–42 [CrossRef]
     [Google Scholar]
  34. Lecoq K., Belloc I., Desgranges C., Konrad M., Daignan-Fornier B. 2001a; YLR209c encodes Saccharomyces cerevisiae purine nucleoside phosphorylase. J Bacteriol 183:4910–4913 [CrossRef]
     [Google Scholar]
  35. Lecoq K., Belloc I., Desgranges C., Daignan-Fornier B. 2001b; Role of adenosine kinase in Saccharomyces cerevisiae : identification of the ADO1 gene and study of the mutant phenotypes. Yeast 18:335–342 [CrossRef]
     [Google Scholar]
  36. Lembo D., Donalisio M., Hofer A., Cornaglia M., Brune W., Koszinowski U., Thelander L., Landolfo S. 2004; The ribonucleotide reductase R1 homolog of murine cytomegalovirus is not a functional enzyme subunit but is required for pathogenesis. J Virol 78:4278–4288 [CrossRef]
     [Google Scholar]
  37. Liu J., Schmitz J. C., Lin X. 6 other authors 2002; Thymidylate synthase as a translational regulator of cellular gene expression. Biochim Biophys Acta 1587174–182 [CrossRef]
     [Google Scholar]
  38. Mao C., Cook W. J., Zhou M., Koszalka G. W., Krenitsky T. A., Ealick S. E. 1997; The crystal structure of Escherichia coli purine nucleoside phosphorylase: a comparison with the human enzyme reveals a conserved topology. Structure 5:1373–1383 [CrossRef]
     [Google Scholar]
  39. McGrogan D. G., Ostland V. E., Byrne P. J., Ferguson H. W. 1998; Systemic disease involving an iridovirus-like agent in cultured tilapia, Oreochromis niloticus L. J Fish Dis 21:149–152 [CrossRef]
     [Google Scholar]
  40. Munagala N., Wang C. C. 2002; The purine nucleoside phosphorylase from Trichomonas vaginalis is a homologue of the bacterial enzyme. Biochemistry 41:10382–10389 [CrossRef]
     [Google Scholar]
  41. Murali S., Wu M.-F., Guo I.-C., Chen S.-C., Yang H.-W., Chang C.-Y. 2002; Molecular characterization and pathogenicity of a grouper iridovirus (GIV) isolated from yellow grouper, Epinephelus awoara (Temminck & Schlegel). J Fish Dis 25:91–100 [CrossRef]
     [Google Scholar]
  42. Németi B., Csanaky I., Gregus Z. 2003; Arsenate reduction in human erythrocytes and rats – testing the role of purine nucleoside phosphorylase. Toxicol Sci 74:22–31 [CrossRef]
     [Google Scholar]
  43. Parker W. B., King S. A., Allan P. W. 8 other authors 1997; In vivo gene therapy of cancer with E. coli purine nucleoside phosphorylase. Hum Gene Ther 8:1637–1644 [CrossRef]
     [Google Scholar]
  44. Pereira H. M., Cleasby A., Pena S. D. J., Franco G. R., Garratt R. C. 2003; Cloning, expression and preliminary crystallographic studies of the potential drug target purine nucleoside phosphorylase from Schistosoma mansoni . Acta Crystallogr D Biol Crystallogr 59:1096–1099 [CrossRef]
     [Google Scholar]
  45. Radabaugh T. R., Sampayo-Reyes A., Zakharyan R. A., Aposhian H. V. 2002; Arsenate reductase II. Purine nucleoside phosphorylase in the presence of dihydrolipoic acid is a route for reduction of arsenate to arsenite in mammalian systems. Chem Res Toxicol 15:692–698 [CrossRef]
     [Google Scholar]
  46. Schnitzler P., Soltau J. B., Fischer M., Reisner H., Scholz J., Delius H., Darai G. 1987; Molecular cloning and physical mapping of the genome of insect iridescent virus type 6: further evidence for circular permutation of the viral genome. Virology 160:66–74 [CrossRef]
     [Google Scholar]
  47. Secrist J. A. III, Parker W. B., Allan P. W. 10 other authors 1999; Gene therapy of cancer: activation of nucleoside prodrugs with E. coli purine nucleoside phosphorylase. Nucleosides Nucleotides 18:745–757 [CrossRef]
     [Google Scholar]
  48. Sudthongkong C., Miyata M., Miyazaki T. 2002; Iridovirus disease in two ornamental tropical freshwater fishes: African lampeye and dwarf gourami. Dis Aquat Organ 48:163–173 [CrossRef]
     [Google Scholar]
  49. Thompson J. D., Gibson T. J., Plewniak F., Jeanmougin F., Higgins D. G. 1997; The clustal_x windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882 [CrossRef]
     [Google Scholar]
  50. Tidona C. A., Darai G. 1997; The complete DNA sequence of lymphocystis disease virus. Virology 230:207–216 [CrossRef]
     [Google Scholar]
  51. Tidona C. A., Darai G. 2000; Iridovirus homologues of cellular genes – implications for the molecular evolution of large DNA viruses. Virus Genes 21:77–81 [CrossRef]
     [Google Scholar]
  52. Tidona C. A., Schnitzler P., Kehm R., Darai G. 1996; Identification of the gene encoding the DNA (cytosine-5) methyltransferase of lymphocystis disease virus. Virus Genes 12:219–229
     [Google Scholar]
  53. Voeks D., Martiniello-Wilks R., Madden V., Smith K., Bennetts E., Both G. W., Russell P. J. 2002; Gene therapy for prostate cancer delivered by ovine adenovirus and mediated by purine nucleoside phosphorylase and fludarabine in mouse models. Gene Ther 9:759–768 [CrossRef]
     [Google Scholar]
  54. Weng S. P., Wang Y. Q., He J. G., Deng M., Lu L., Guan H. J., Liu Y. J., Chan S.-M. 2002; Outbreaks of an iridovirus in red drum, Sciaenops ocellata (L.), cultured in southern China. J Fish Dis 25:681–685 [CrossRef]
     [Google Scholar]
  55. Whittington R. J., Philbey A., Reddacliff G. L., MacGown A. R. 1994; Epidemiology of epizootic haematopoietic necrosis virus (EHNV) infection in farmed rainbow trout, Oncorhynchus mykiss (Walbaum): findings based on virus isolation, antigen capture ELISA and serology. J Fish Dis 17:205–218 [CrossRef]
     [Google Scholar]
  56. Williams T. 1996; The iridoviruses. Adv Virus Res 46:345–412
     [Google Scholar]
  57. Williams S. R., Goddard J. M., Martin D. W. Jr 1984; Human purine nucleoside phosphorylase cDNA sequence and genomic clone characterization. Nucleic Acids Res 12:5779–5787 [CrossRef]
     [Google Scholar]
  58. Williams T., Chinchar G., Darai G., Hyatt A., Kalmakoff J., Seligy V. 2000; Iridoviridae . In Virus Taxonomy: Seventh Report of the International Committee on Taxonomy of Viruses . pp  167–182 Edited by van Regenmortel M. H. V., Fauquet C. M., Bishop D. H. L., Carstens E. B., Estes M. K., Lemon S. M., Maniloff J., Mayo M. A., McGeoch D. J., Pringle C. R., Wickner R. B. San Diego: Academic Press;
  59. Willis D. B., Granoff A. 1980; Frog virus 3 DNA is heavily methylated at CpG sequences. Virology 107:250–257 [CrossRef]
     [Google Scholar]
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Identification and characterization of a novel gene of grouper iridovirus encoding a purine nucleoside phosphorylase
J. Gen. Virol. 85, 2883 (2004); https://doi.org/10.1099/vir.0.80249-0
/content/journal/jgv/10.1099/vir.0.80249-0
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