1887

Journal of General Virology header logo

Volume 75, Issue 12

Localization of Bunyamwera bunyavirus G1 glycoprotein to the Golgi requires association with G2 but not with NSm Free

Abstract

The Bunyamwera bunyavirus (BUN) M RNA genome segment encodes three proteins, two glycoproteins termed G1 and G2 and a non-structural protein called NSm, in the form of a polyprotein precursor that is co-translationally cleaved to give the mature proteins. Indirect immunofluorescence experiments have shown that these proteins localize to the Golgi complex in BUN-infected cells. We have used a recombinant vaccinia virus (vTF7-3), which expresses bacteriophage T7 RNA polymerase, to drive the expression of plasmids containing either the entire BUN M segment cDNA or fragments that encode the G1, G2 and NSm proteins separately under control of the T7 promoter. After transfection of these plasmids into vTF7-3-infected cells, correctly sized and processed proteins were detected by immunoprecipitation with BUN-specific antibodies. Immunofluorescence experiments showed that G1, G2 and NSm localized to the Golgi when transiently expressed from the full-length cDNA. When G2 or NSm were expressed separately they also localized to the Golgi, but when G1 was expressed alone a staining pattern typical for the endoplasmic reticulum was obtained. However coexpression of G2 and G1 from independent plasmids resulted in G1 localizing to the Golgi. In contrast translocation of G1 to the Golgi was not observed when G1 was coexpressed with NSm, although NSm itself was still detected in the Golgi. Similar results were obtained when the proteins were expressed from transfected plasmids containing the G2-, NSm- or G1-coding sequences under control of the cytomegalovirus immediate-early promoter. The localization of G1 to the Golgi when coexpressed with G2 was confirmed by the loss of endoglycosidase H (endo H) sensitivity of G1 after approximately 60 min in a pulse-chase experiment; G1 remained sensitive to endo H when expressed either alone or in combination with NSm. These results suggest that G2 contains the Golgi targeting and/or retention signals and that G1 has to interact with this protein to localize to this cellular compartment.

  • Received:
  • Accepted:
  • Published Online:
© The Journal of General Virology 1994
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-75-12-3441
1994-12-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/75/12/JV0750123441.html?itemId=/content/journal/jgv/10.1099/0022-1317-75-12-3441&mimeType=html&fmt=ahah

References

  1. Bishop D. H. L. 1990; Bunyaviridae and their replication. I. Bunyaviridae. In Virology, 2nd edn. pp 1155–1173 Fields B. N., Knipe D. M. Edited by New York: Raven Press;
     [Google Scholar]
  2. Chen S.-Y., Matsuoka Y., Compans R. W. 1991; Golgi complex localisation of the Punta Toro virus G2 protein requires its association with the G1 protein. Virology 183:351–365
     [Google Scholar]
  3. Elliott R. M. 1990; Review article. Molecular biology of the Bunyaviridae. Journal of General Virology 71:501–522
     [Google Scholar]
  4. Elliott R. M., Schmaljohn C. S., Collett M. S. 1991; Bunyaviridae genome structure and gene expression. Current Topics in Microbiology and Immunology 169:91–141
     [Google Scholar]
  5. Fazakerley J. K., Gonzalez-Scarano F., Strickler J., Dietzschold B., Karush F., Nathanson N. 1988; Organization of the middle RNA segment of snowshoe hare bunyavirus. Virology 167:422–432
     [Google Scholar]
  6. Fuerst T. F., Niles E. G., Studier F. W., Moss B. 1986; Eukaryotic expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proceedings of the National Academy of Sciences U.S.A.: 838122–8126
     [Google Scholar]
  7. Fuerst T. F., Earl P. L., Moss B. 1987; Use of a hybrid vaccinia virus-T7 RNA polymerase system for expression of target genes. Molecular and Cellular Biology 7:2538–2544
     [Google Scholar]
  8. Gonzalez-Scarano F., Shore R. E., Calisher C. H., Nathanson N. 1982; Characterization of monoclonal antibodies against the G1 and N protein of La Crosse and Tahyna, two California serogroup bunyaviruses. Virology 120:42–53
     [Google Scholar]
  9. Ihara T., Smith J., Dalrymple J. M., Bishop D. H. L. 1985; Complete sequence of the glycoproteins and M RNA of Punta Toro phlebovirus compared to those of Rift Valley fever virus. Virology 144:246–259
     [Google Scholar]
  10. Kakach L. T., Wasmoen T. L., Collett M. S. 1988; Rift Valley fever virus M segment: use of recombinant vaccinia viruses to study Phlebovirusgene expression. Journal of Virology 62:826–833
     [Google Scholar]
  11. Kornfeld R., Kornfeld S. 1985; Assembly of asparagine-linked oligosaccharides. Annual Review of Biochemistry 54:631–644
     [Google Scholar]
  12. Law M. D., Speck J., Moyer J. W. 1992; The M RNA of Impatiens necrotic spot Tospovirus (Bunyaviridae) has an ambisense genomic organization. Virology 188:732–741
     [Google Scholar]
  13. Lees J. F., Pringle C. R., Elliott R. M. 1986; Nucleotide sequence of the Bunyamwera virus M RNA segment: conservation of structural features in the bunyavirus glycoprotein gene product. Virology 148:1–14
     [Google Scholar]
  14. Machamer C. E. 1991; Golgi retention signals: do membranes hold the key?. Trends in Cell Biology 1:141–144
     [Google Scholar]
  15. Mackett M., Smith G. L., Moss B. 1985; The construction and characterization of vaccinia virus recombinants expressing foreign genes. In DNA Cloning 2 pp 191–211 Glover D. M. Edited by Oxford: IRL Press;
     [Google Scholar]
  16. Maddon P. J., Dalgleish A. G., McDougal J. S., Clapham P. R., Weiss R. A., Axel R. 1986; The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain. Cell 47:333–348
     [Google Scholar]
  17. Marriott A. C., El-Ghorr A. A., Nuttall P. A. 1992; Dugbe nairovirus M RNA: nucleotide sequence and coding strategy. Virology 190:606–615
     [Google Scholar]
  18. Matsuoka Y., Ihara T., Bishop D. H. L., Compans R. W. 1988; Intracellular accumulation of Punta Toro virus glycoproteins expressed from cloned cDNA. Virology 167:251–267
     [Google Scholar]
  19. Matsuoka Y., Chen S.-Y., Compans R. W. 1991; Bunyavirus protein transport and assembly. Current Topics in Microbiology and Immunology 169:161–179
     [Google Scholar]
  20. Munro S. 1991; Sequences within and adjacent to the transmembrane segment of α-2,6-sialyltransferase specify Golgi retention. EMBO Journal 10:3577–3588
     [Google Scholar]
  21. Nakitare G. W. 1992; Expression of the Bunyamwera virus M genome segment gene products. Ph.D. Thesis University of Glasgow:
     [Google Scholar]
  22. Nakitare G. W., Elliott R. M. 1993; Expression of the Bunyamwera virus M genome segment and intracellular localisation of NSm. Virology 195:511–520
     [Google Scholar]
  23. Nilsson T., Lucocq J. M., Mackay D., Warren G. 1991; The membrane spanning domain of β-l,4-galactosyltransferase specifies transGolgi localisation. EMBO Journal 10:3567–3575
     [Google Scholar]
  24. Pensiero M. N., Hay J. 1992; The Hantaan virus M-segment glycoproteins G1 and G2 can be expressed independently. Journal of Virology 66:1907–1914
     [Google Scholar]
  25. Rönnholm R. 1992; Localization to the Golgi complex of Uukuniemi virus glycoproteins G1 and G2 expressed from cloned cDNAs. Journal of Virology 66:4525–1531
     [Google Scholar]
  26. Rönnholm R., Pettersson R. F. 1987; Complete nucleotide sequence of the M RNA segment of Uukuniemi virus encoding the membrane glycoproteins G1 and G2. Virology 160:191–202
     [Google Scholar]
  27. Rose J. K., Buonocore L., Whitt M. A. 1991; A new cationic liposome reagent mediating nearly quantitative transfection of animal cells. Biotechniques 10:520–525
     [Google Scholar]
  28. Ruusala A., Persson R., Schmaljohn C. S., Pettersson R. F. 1992; Coexpression of the membrane glycoproteins G1 and G2 of Hantaan virus is required for targetting to the Golgi complex. Virology 186:53–64
     [Google Scholar]
  29. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  30. Schmaljohn C. S., Patterson J. L. 1990; Bunyaviridae and their replication. II. Replication of Bunyaviridae. In Virology, 2nd edn.. pp 1175–1194 Fields B. N., Knipe D. M. Edited by New York: Raven Press;
     [Google Scholar]
  31. Schmaljohn C. S., Schmaljohn A. L., Dalrymple J. M. 1987; Hantaan virus M RNA: coding strategy, nucleotide sequence and gene order. Virology 157:31–39
     [Google Scholar]
  32. Stow N. D., Hammarste O., Arbuckle M. I., Elias P. 1993; Inhibition of herpes simplex virus type 1 DNA replication by mutant forms of the origin-binding protein. Virology 196:413–418
     [Google Scholar]
  33. Swift A. M., Machamer C. E. 1991; A Golgi retention signal in a membrane-spanning domain of coronavirus El protein. Journal of Cell Biology 115:19–30
     [Google Scholar]
  34. Watret G. E., Pringle C. R., Elliott R. M. 1985; Synthesis of bunyavirus-specific proteins in a continuous cell line (XTC-2) derived from Xenopus laevis. Journal of General Virology 66:473–482
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-75-12-3441
Loading
Localization of Bunyamwera bunyavirus G1 glycoprotein to the Golgi requires association with G2 but not with NSm
J. Gen. Virol. 75, 3441 (1994); https://doi.org/10.1099/0022-1317-75-12-3441
/content/journal/jgv/10.1099/0022-1317-75-12-3441
/content/journal/jgv/10.1099/0022-1317-75-12-3441
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error