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. 1992 Jul;66(7):4525–4531. doi: 10.1128/jvi.66.7.4525-4531.1992

Localization to the Golgi complex of Uukuniemi virus glycoproteins G1 and G2 expressed from cloned cDNAs.

R Rönnholm 1
PMCID: PMC241262  PMID: 1602557

Abstract

The membrane glycoproteins G1 and G2 of Uukuniemi virus, a bunyavirus, accumulate in the Golgi complex (GC) during virus infection. These proteins have therefore been considered to be good models for studying the intracellular transport to and retention in the GC. In this study, I have used indirect immunofluorescence to localize in COS cells the Uukuniemi virus glycoproteins G1 and G2 expressed together or separately from cloned cDNAs with use of simian virus 40-based vectors. When expressed together from the full-length cDNA, G1 and G2 were correctly translocated, processed, and targeted to the GC, indicating that the information for GC targeting resides in the proteins. When the proteins were expressed separately, G1 was transported to the GC and retained there. In contrast, G2 could not be detected in the GC but was most probably retained and finally degraded in the endoplasmic reticulum. However, in cells cotransfected with G1 and G2 cDNAs, the proteins could both again be found in the GC. These results suggest that G1 is a responsible for targeting to and retention of the Uukuniemi virus glycoproteins in the GC. G2 would thus accumulate in the GC by virtue of its binding to G1.

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Selected References

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  1. Braakman I., Hoover-Litty H., Wagner K. R., Helenius A. Folding of influenza hemagglutinin in the endoplasmic reticulum. J Cell Biol. 1991 Aug;114(3):401–411. doi: 10.1083/jcb.114.3.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen S. Y., Compans R. W. Oligomerization, transport, and Golgi retention of Punta Toro virus glycoproteins. J Virol. 1991 Nov;65(11):5902–5909. doi: 10.1128/jvi.65.11.5902-5909.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen S. Y., Matsuoka Y., Compans R. W. Assembly and polarized release of Punta Toro virus and effects of brefeldin A. J Virol. 1991 Mar;65(3):1427–1439. doi: 10.1128/jvi.65.3.1427-1439.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen S. Y., Matsuoka Y., Compans R. W. Golgi complex localization of the Punta Toro virus G2 protein requires its association with the G1 protein. Virology. 1991 Jul;183(1):351–365. doi: 10.1016/0042-6822(91)90148-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gahmberg N., Kuismanen E., Keränen S., Pettersson R. F. Uukuniemi virus glycoproteins accumulate in and cause morphological changes of the Golgi complex in the absence of virus maturation. J Virol. 1986 Mar;57(3):899–906. doi: 10.1128/jvi.57.3.899-906.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hurtley S. M., Helenius A. Protein oligomerization in the endoplasmic reticulum. Annu Rev Cell Biol. 1989;5:277–307. doi: 10.1146/annurev.cb.05.110189.001425. [DOI] [PubMed] [Google Scholar]
  7. Jackson M. R., Nilsson T., Peterson P. A. Identification of a consensus motif for retention of transmembrane proteins in the endoplasmic reticulum. EMBO J. 1990 Oct;9(10):3153–3162. doi: 10.1002/j.1460-2075.1990.tb07513.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jasmin B. J., Cartaud J., Bornens M., Changeux J. P. Golgi apparatus in chick skeletal muscle: changes in its distribution during end plate development and after denervation. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7218–7222. doi: 10.1073/pnas.86.18.7218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Joyce G. F., Inoue T. A novel technique for the rapid preparation of mutant RNAs. Nucleic Acids Res. 1989 Jan 25;17(2):711–722. [PMC free article] [PubMed] [Google Scholar]
  10. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kuismanen E., Bång B., Hurme M., Pettersson R. F. Uukuniemi virus maturation: immunofluorescence microscopy with monoclonal glycoprotein-specific antibodies. J Virol. 1984 Jul;51(1):137–146. doi: 10.1128/jvi.51.1.137-146.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kuismanen E., Hedman K., Saraste J., Pettersson R. F. Uukuniemi virus maturation: accumulation of virus particles and viral antigens in the Golgi complex. Mol Cell Biol. 1982 Nov;2(11):1444–1458. doi: 10.1128/mcb.2.11.1444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kuismanen E. Posttranslational processing of Uukuniemi virus glycoproteins G1 and G2. J Virol. 1984 Sep;51(3):806–812. doi: 10.1128/jvi.51.3.806-812.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kuismanen E., Saraste J., Pettersson R. F. Effect of monensin on the assembly of Uukuniemi virus in the Golgi complex. J Virol. 1985 Sep;55(3):813–822. doi: 10.1128/jvi.55.3.813-822.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Matsuoka Y., Ihara T., Bishop D. H., Compans R. W. Intracellular accumulation of Punta Toro virus glycoproteins expressed from cloned cDNA. Virology. 1988 Nov;167(1):251–260. doi: 10.1016/0042-6822(88)90075-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maxwell I. H., Maxwell F. Electroporation of mammalian cells with a firefly luciferase expression plasmid: kinetics of transient expression differ markedly among cell types. DNA. 1988 Oct;7(8):557–562. doi: 10.1089/dna.1.1988.7.557. [DOI] [PubMed] [Google Scholar]
  17. Pensiero M. N., Jennings G. B., Schmaljohn C. S., Hay J. Expression of the Hantaan virus M genome segment by using a vaccinia virus recombinant. J Virol. 1988 Mar;62(3):696–702. doi: 10.1128/jvi.62.3.696-702.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Persson R., Pettersson R. F. Formation and intracellular transport of a heterodimeric viral spike protein complex. J Cell Biol. 1991 Jan;112(2):257–266. doi: 10.1083/jcb.112.2.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pesonen M., Kuismanen E., Pettersson R. F. Monosaccharide sequence of protein-bound glycans of Uukuniemi virus. J Virol. 1982 Feb;41(2):390–400. doi: 10.1128/jvi.41.2.390-400.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pettersson R. F., Hewlett M. J., Baltimore D., Coffin J. M. The genome of Uukuniemi virus consists of three unique RNA segments. Cell. 1977 May;11(1):51–63. doi: 10.1016/0092-8674(77)90316-6. [DOI] [PubMed] [Google Scholar]
  21. Pettersson R. F. Protein localization and virus assembly at intracellular membranes. Curr Top Microbiol Immunol. 1991;170:67–106. doi: 10.1007/978-3-642-76389-2_3. [DOI] [PubMed] [Google Scholar]
  22. Ranki M., Pettersson R. F. Uukuniemi virus contains an RNA polymerase. J Virol. 1975 Dec;16(6):1420–1425. doi: 10.1128/jvi.16.6.1420-1425.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rose J. K., Doms R. W. Regulation of protein export from the endoplasmic reticulum. Annu Rev Cell Biol. 1988;4:257–288. doi: 10.1146/annurev.cb.04.110188.001353. [DOI] [PubMed] [Google Scholar]
  24. Rottier P. J., Rose J. K. Coronavirus E1 glycoprotein expressed from cloned cDNA localizes in the Golgi region. J Virol. 1987 Jun;61(6):2042–2045. doi: 10.1128/jvi.61.6.2042-2045.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ruusala A., Persson R., Schmaljohn C. S., Pettersson R. F. Coexpression of the membrane glycoproteins G1 and G2 of Hantaan virus is required for targeting to the Golgi complex. Virology. 1992 Jan;186(1):53–64. doi: 10.1016/0042-6822(92)90060-3. [DOI] [PubMed] [Google Scholar]
  26. Rönnholm R., Pettersson R. F. Complete nucleotide sequence of the M RNA segment of Uukuniemi virus encoding the membrane glycoproteins G1 and G2. Virology. 1987 Sep;160(1):191–202. doi: 10.1016/0042-6822(87)90060-2. [DOI] [PubMed] [Google Scholar]
  27. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  28. Shin J., Dunbrack R. L., Jr, Lee S., Strominger J. L. Signals for retention of transmembrane proteins in the endoplasmic reticulum studied with CD4 truncation mutants. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1918–1922. doi: 10.1073/pnas.88.5.1918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ulmanen I., Seppälä P., Pettersson R. F. In vitro translation of Uukuniemi virus-specific RNAs: identification of a nonstructural protein and a precursor to the membrane glycoproteins. J Virol. 1981 Jan;37(1):72–79. doi: 10.1128/jvi.37.1.72-79.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wasmoen T. L., Kakach L. T., Collett M. S. Rift Valley fever virus M segment: cellular localization of M segment-encoded proteins. Virology. 1988 Sep;166(1):275–280. doi: 10.1016/0042-6822(88)90174-2. [DOI] [PubMed] [Google Scholar]
  31. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol. 1983;100:468–500. doi: 10.1016/0076-6879(83)00074-9. [DOI] [PubMed] [Google Scholar]

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