1887

Abstract

The nucleoprotein (NP) of Marburg virus (MBG), a filovirus, is encoded by the gene closest to the 3′ end of the non-segmented negative-strand RNA genome. Sequence comparison has indicated that NP is the functional equivalent to the nucleoproteins of paramyxoviruses and rhabdoviruses. Expression of recombinant NP in two eukaryotic systems using vaccinia virus and baculovirus (vectors pSCll and pAcYMBl, respectively) and analysis of MBG-specific proteins have demonstrated that the NP of MBG is phosphorylated. The NP appeared in two forms differing in by about 2K (94K and 92K respectively). Dephosphorylation clearly demonstrated that the 94K form is phosphorylated whereas the 92K form is unphosphorylated. In virion particles NP was exclusively present in the phosphorylated form. These findings suggest that only the phosphorylated NP can form nucleocapsid complexes and interact with the genomic RNA.

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1994-04-01
2022-08-17
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References

  1. Banerjee A. K., Barik S. 1992; Gene expression of vesicular stomatitis virus genome RNA. Virology 188:417–128
    [Google Scholar]
  2. Barr J., Chambers P., Pringle C. R., Easton A. J. 1991; Sequence of the major nucleocapsid protein gene of pneumonia virus of mice: sequence comparisons suggest structural homology between nucleocapsid proteins of pneumoviruses, paramyxoviruses, rhabdo- viruses and filoviruses. Journal of General Virology 72:677–685
    [Google Scholar]
  3. Becker S., Feldmann H., Will C., Slenczka W. 1992; Evidence for filovirus antibodies in humans and imported monkeys: do subclinical filovirus infections occur worldwide?. Medical Microbiology and Immunology 181:43–55
    [Google Scholar]
  4. Bressan G., Stanley K. 1987; pUEX, a bacterial expression vector related to pEX with universal host specificity. Nucleic Acids Research 15:10056
    [Google Scholar]
  5. Chakrabarti S., Brechling K., Moss B. 1985; Vaccinia virus expression vector: coexpression of β-galactosidase provides visual screening of recombinant virus plaques. Molecular and Cellular Biology 5:3403–3409
    [Google Scholar]
  6. Cohen P. 1985; The role of protein phosphorylation in the hormone control of enzyme activity. European Journal of Biochemistry 151:439–148
    [Google Scholar]
  7. Einberger H., Mertz R., Hofschneider P. H., Neubert W. J. 1990; Purification, renaturation and reconstituted protein kinase activity of the Sendai virus large (L) protein: L protein phosphoryl- ates the NP and P protein in vitro . Journal of Virology 64:4274–4280
    [Google Scholar]
  8. Elliott L., Kiley M. P., McCormick J. B. 1985; Descriptive analysis of Ebola virus proteins. Virology 147:169–176
    [Google Scholar]
  9. Feldmann H., Mühlberger E., Randolf A., Will C., Kiley M. P., Sanchez A., Klenk H.-D. 1992; Marburg virus, a filovirus: messenger RNAs, gene order, and regulatory elements of the replication cycle. Virus Research 24:1–19
    [Google Scholar]
  10. Felgner P. L., Gadek T. R., Fiolm M., Roman R., Chan FI. W., Wenz M., Northrop J. P., Ringold G. M., Danielsen M. 1987; Lipofection: a highly efficient, lipid-mediated DNA- transfection procedure. Proceedings of the National Academy of Sciences U.S.A: 847413–7417
    [Google Scholar]
  11. Grässer F. A., König S. 1992; Phosphorylation of SV40 large T antigen at threonine residues results in conversion to a lower apparent molecular weight form. Archives of Virology 126:313–320
    [Google Scholar]
  12. Green P. L., Yip M. T., Xie Y., Chen I. S. Y. 1992; Phosphorylation regulates RNA binding by the human T-cell leukemia virus rex protein. Journal of Virology 66:4325–1330
    [Google Scholar]
  13. Han X.-M., Laras A., Rounseville M. P., Kumar A., Shank P. R. 1992; Human immunodeficiency virus type 1 tat-mediated trans activation correlates with the phosphorylation state of a cellular tar stem binding factor. Journal of Virology 66:4065–4072
    [Google Scholar]
  14. Huber M., Cattaneo R., Spielhofer P., Ürvell C., Norrby E., Messerli M., Perriard J.-C., Billeter M. A. 1991; Measles virus phosphoprotein retains the nucleocapsid protein in the cytoplasm. Virology 185:299–308
    [Google Scholar]
  15. Kiley M. P., Bowen E. T. W., Eddy G. A., Isaäcson M., Johnson K. M., Mccormick J. B., Murphy F. A., Pattyn S. R., Peters D., Prozesky O. W., Regnery R. L., Simpson D. I. H., Slenczka W., Sureau P., Vandergroen G., Webb P. A., Wulff H. 1982; Filovmdae: a taxonomic home for Marburg and Ebola viruses?. Intervirology 18:24–32
    [Google Scholar]
  16. Kiley M. P., Cox N. J., Elliott L. H., Sanchez A., DeFries R., Buchmeier M. J., Richman D. D., Mccormick J. B. 1988; Physicochemical properties of Marburg virus: evidence for three distinct virus strains and their relationship to Ebola virus. Journal of General Virology 69:1957–1967
    [Google Scholar]
  17. Kingsbury D. W. editor 1991 The Paramyxoviruses New York & London: Plenum Press;
    [Google Scholar]
  18. Kretzschmar E., Veit M., Brunschön S., Kuroda K., Klenk H.-D. 1992; Secretion of fowl plague virus haemagglutinin from insect cells requires elimination of both hydrophobic domains. Journal of General Virology 73:839–848
    [Google Scholar]
  19. Kuroda K., Hauser C., Rott R., Klenk H.-D., Doerfler W. 1986; Expression of the influenza virus haemagglutinin in insect cells by a baculovirus vector. EM BO Journal 5:1359–1365
    [Google Scholar]
  20. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 111680–685
    [Google Scholar]
  21. Lamb R. A., Choppin P. W. 1977; The synthesis of Sendai virus polypeptides. III. Phosphorylation of polypeptides. Virology 81:382–397
    [Google Scholar]
  22. Leader D. P., Katan M. 1988; Viral aspects of protein phosphorylation. Journal of General Virology 69:1441–1464
    [Google Scholar]
  23. Luckow V. A., Summers M. D. 1988; Trends in the development of baculovirus expression vectors. Bio/Technology 6:47–55
    [Google Scholar]
  24. Mackett M., Smith G. L., Moss B. 1986; The construction and characterization of vaccinia virus recombinants expressing foreign genes. In DNA Cloning: A Practical Approach 2 pp 191–211 Glover D. M. Edited by Oxford: IRL Press;
    [Google Scholar]
  25. Matsuura Y., Possee R., Overton H. A., Bishop D. H. L. 1987; Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. Journal of General Virology 68:1233–1250
    [Google Scholar]
  26. Morgan E. M. 1991; Evolutionary relationships of paramyxovirus nucleocapsid-associated proteins. In The Paramyxoviruses pp 163–179 Kingsbury D. W. Edited by New York & London: Plenum Press;
    [Google Scholar]
  27. Mühlberger E., Sanchez A., Randolf A., Will C., Klenk H.-D., Feldmann H. 1992; The nucleotide sequence of the L gene of Marburg virus, a filovirus: homologies with paramyxoviruses and rhabdoviruses. Virology 187:534–547
    [Google Scholar]
  28. Murphy F. A., Kiley M. P., Fisher-Hoch S. P. 1990; Filoviridae: Marburg and Ebola viruses. In Virology, 2nd edn. pp 933–944 Fields B. N., Knipe D. M. Edited by New York: Raven Press;
    [Google Scholar]
  29. Nichol S. T. 1993; Vesicular stomatitis virus New Jersey and Indiana. In Encyclopedia of Virology Webster R. G., Granoff A. Edited by London: Saunders Scientific Publications; in press
    [Google Scholar]
  30. Norrby E., Oxman M. N. 1990; Measles virus. In Virology, 2nd edn.. pp 1013–1044 Fields B. N., Knipe D. M. Edited by New York: Raven Press;
    [Google Scholar]
  31. Pen J., Welling G. W., Welling-Wester S. 1989; An efficient procedure for the isolation of recombinant baculovirus. Nucleic Acids Research 17:451
    [Google Scholar]
  32. Préaud C., Harris R. D., Fulop V., Koh C.-L., Wong J., Flamand A., Bishop D. H. L. 1990; Expression, characterization, and purification of a phosphorylated rabies nucleoprotein synthesized in insect cells by baculovirus vectors. Virology 178:486–497
    [Google Scholar]
  33. Pringle C. R. 1991; The order Mononegavirales. Archives of Virology 117:137–140
    [Google Scholar]
  34. Rhim J. S., Cho H. Y., Huebner R. J. 1975; Non-producer human cells induced by murine sarcoma virus. International Journal of Cancer 15:23–29
    [Google Scholar]
  35. Rohrmann G., Yeun L., Moss B. 1986; Transcription of vaccinia virus early genes by enzymes isolated from vaccinia virions terminates downstream of a regulatory sequence. Cell 46:1029–1035
    [Google Scholar]
  36. Sanchez A., Kiley M. P., Klenk H.-D., Feldmann H. 1992; Sequence analysis of the Marburg virus nucleoprotein gene: comparison to Ebola virus and other non-segmented negative-strand RNA viruses. Journal of General Virology 73:347–357
    [Google Scholar]
  37. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences U.S.A.: 745463–5467
    [Google Scholar]
  38. Schnittler H.-J., Mahner F., Drenckhahn D., Klenk H.-D., Feldmann H. 1993; Replication of Marburg virus in human endothelial cells: a possible mechanism for the development of viral hemorrhagic disease. Journal of Clinical Investigation 91:1301–1309
    [Google Scholar]
  39. Smith D. H., Johnson B. K., Isaäcson M., Swanapoel R., Johnson K. M., Kiley M. P., Bagshawe A., Siongok T., Keruga W. K. 1982; Marburg-virus disease in Kenya. Lancet i:816–820
    [Google Scholar]
  40. Sokol F., Clark H. F. 1973; Phosphoproteins, structural components of rhabdoviruses. Virology 52:246–263
    [Google Scholar]
  41. Spehner D., Kirn A., Drillien R. 1991; Assembly of nucleocapsid-like structures in animal cells infected with a vaccinia virus recombinant encoding the measles virus nucleoprotein. Journal of Virology 65:6296–6300
    [Google Scholar]
  42. Sprague J., Condra J. H., Arnheiter H., Lazzarini R. A. 1983; Expression of a recombinant DNA gene coding for the vesicular stomatitis virus nucleocapsid protein. Journal of Virology 45:773–778
    [Google Scholar]
  43. Wagner R. R. 1987 The Rhabdoviruses New York & London: Plenum Press;
    [Google Scholar]
  44. Will C., Mühlberger E., Linder D., Slenczka W., Klenk H.-D., Feldmann H. 1993; Marburg virus gene four encodes the virion membrane protein, a type I transmembrane glycoprotein. Journal of Virology 67:1203–1210
    [Google Scholar]
  45. Zhirnov O. P., Bukrinskaya A. G. 1981; Two forms of influenza virus nucleoprotein in infected cells and virions. Virology 109:174–179
    [Google Scholar]
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