Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Sep;87(18):6944–6948. doi: 10.1073/pnas.87.18.6944

The E1 glycoprotein of an avian coronavirus is targeted to the cis Golgi complex.

C E Machamer 1, S A Mentone 1, J K Rose 1, M G Farquhar 1
PMCID: PMC54658  PMID: 2169615

Abstract

It was previously reported that the E1 protein of an avian coronavirus was targeted to the juxtanuclear region in COS cells expressing the protein from cloned cDNA, suggesting that the protein contains information for targeting to the Golgi complex. The first of three membrane-spanning domains was required for intracellular targeting, because a mutant E1 (delta m1,2) lacking this domain was delivered to the plasma membrane. We have used immunoelectron microscopy to localize the wild-type E1 protein within Golgi elements of COS cells and AtT-20 cells expressing these proteins from recombinant vaccinia vectors. By immunoperoxidase and immunogold labeling, the wild-type E1 protein was localized to one or two cisternae located on one side of the Golgi stack that could be identified as the cis side in AtT-20 cells. In contrast, the mutant E1 protein was detected in all cisternae across the stack as well as at the plasma membrane. When the E1 proteins were immunoprecipitated and subjected to digestion with endoglycosidase H, the majority of the wild-type E1 glycoprotein was endoglycosidase H sensitive, whereas the majority of the mutant E1 was processed to an endoglycosidase H-resistant, polylactosaminoglycan-containing form. The findings indicate that the wild-type E1 protein is specifically targeted to cis Golgi cisternae and are consistent with the assumption that the first membrane-spanning domain is required for targeting to the cis Golgi.

Full text

PDF
6944

Images in this article

6945Image
on p.6945
6946Image
on p.6946
6947Image
on p.6947
6948Image
on p.6948

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Armstrong J., McCrae M., Colman A. Expression of coronavirus E1 and rotavirus VP10 membrane proteins from synthetic RNA. J Cell Biochem. 1987 Oct;35(2):129–136. doi: 10.1002/jcb.240350206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boursnell M. E., Brown T. D., Binns M. M. Sequence of the membrane protein gene from avian coronavirus IBV. Virus Res. 1984;1(4):303–313. doi: 10.1016/0168-1702(84)90019-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown W. J., Constantinescu E., Farquhar M. G. Redistribution of mannose-6-phosphate receptors induced by tunicamycin and chloroquine. J Cell Biol. 1984 Jul;99(1 Pt 1):320–326. doi: 10.1083/jcb.99.1.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cummings R. D., Kornfeld S. The distribution of repeating [Gal beta 1,4GlcNAc beta 1,3] sequences in asparagine-linked oligosaccharides of the mouse lymphoma cell lines BW5147 and PHAR 2.1. J Biol Chem. 1984 May 25;259(10):6253–6260. [PubMed] [Google Scholar]
  5. D'Agostaro G., Bendiak B., Tropak M. Cloning of cDNA encoding the membrane-bound form of bovine beta 1,4-galactosyltransferase. Eur J Biochem. 1989 Jul 15;183(1):211–217. doi: 10.1111/j.1432-1033.1989.tb14915.x. [DOI] [PubMed] [Google Scholar]
  6. Dunphy W. G., Rothman J. E. Compartmental organization of the Golgi stack. Cell. 1985 Aug;42(1):13–21. doi: 10.1016/s0092-8674(85)80097-0. [DOI] [PubMed] [Google Scholar]
  7. Farquhar M. G., Palade G. E. The Golgi apparatus (complex)-(1954-1981)-from artifact to center stage. J Cell Biol. 1981 Dec;91(3 Pt 2):77s–103s. doi: 10.1083/jcb.91.3.77s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Farquhar M. G. Progress in unraveling pathways of Golgi traffic. Annu Rev Cell Biol. 1985;1:447–488. doi: 10.1146/annurev.cb.01.110185.002311. [DOI] [PubMed] [Google Scholar]
  9. Fukuda M. Cell surface glycoconjugates as onco-differentiation markers in hematopoietic cells. Biochim Biophys Acta. 1985;780(2):119–150. doi: 10.1016/0304-419x(84)90002-7. [DOI] [PubMed] [Google Scholar]
  10. Fukuda M., Guan J. L., Rose J. K. A membrane-anchored form but not the secretory form of human chorionic gonadotropin-alpha chain acquires polylactosaminoglycan. J Biol Chem. 1988 Apr 15;263(11):5314–5318. [PubMed] [Google Scholar]
  11. Joziasse D. H., Shaper J. H., Van den Eijnden D. H., Van Tunen A. J., Shaper N. L. Bovine alpha 1----3-galactosyltransferase: isolation and characterization of a cDNA clone. Identification of homologous sequences in human genomic DNA. J Biol Chem. 1989 Aug 25;264(24):14290–14297. [PubMed] [Google Scholar]
  12. Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
  13. Larsen R. D., Rajan V. P., Ruff M. M., Kukowska-Latallo J., Cummings R. D., Lowe J. B. Isolation of a cDNA encoding a murine UDPgalactose:beta-D-galactosyl- 1,4-N-acetyl-D-glucosaminide alpha-1,3-galactosyltransferase: expression cloning by gene transfer. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8227–8231. doi: 10.1073/pnas.86.21.8227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Machamer C. E., Rose J. K. A specific transmembrane domain of a coronavirus E1 glycoprotein is required for its retention in the Golgi region. J Cell Biol. 1987 Sep;105(3):1205–1214. doi: 10.1083/jcb.105.3.1205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Masri K. A., Appert H. E., Fukuda M. N. Identification of the full-length coding sequence for human galactosyltransferase (beta-N-acetylglucosaminide: beta 1,4-galactosyltransferase). Biochem Biophys Res Commun. 1988 Dec 15;157(2):657–663. doi: 10.1016/s0006-291x(88)80300-0. [DOI] [PubMed] [Google Scholar]
  16. Mayer T., Tamura T., Falk M., Niemann H. Membrane integration and intracellular transport of the coronavirus glycoprotein E1, a class III membrane glycoprotein. J Biol Chem. 1988 Oct 15;263(29):14956–14963. doi: 10.1016/S0021-9258(18)68131-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Merkle R. K., Cummings R. D. Relationship of the terminal sequences to the length of poly-N-acetyllactosamine chains in asparagine-linked oligosaccharides from the mouse lymphoma cell line BW5147. Immobilized tomato lectin interacts with high affinity with glycopeptides containing long poly-N-acetyllactosamine chains. J Biol Chem. 1987 Jun 15;262(17):8179–8189. [PubMed] [Google Scholar]
  18. Moremen K. W. Isolation of a rat liver Golgi mannosidase II clone by mixed oligonucleotide-primed amplification of cDNA. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5276–5280. doi: 10.1073/pnas.86.14.5276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakazawa K., Ando T., Kimura T., Narimatsu H. Cloning and sequencing of a full-length cDNA of mouse N-acetylglucosamine (beta 1-4)galactosyltransferase. J Biochem. 1988 Aug;104(2):165–168. doi: 10.1093/oxfordjournals.jbchem.a122434. [DOI] [PubMed] [Google Scholar]
  20. Rajan V. P., Larsen R. D., Ajmera S., Ernst L. K., Lowe J. B. A cloned human DNA restriction fragment determines expression of a GDP-L-fucose: beta-D-galactoside 2-alpha-L-fucosyltransferase in transfected cells. Evidence for isolation and transfer of the human H blood group locus. J Biol Chem. 1989 Jul 5;264(19):11158–11167. [PubMed] [Google Scholar]
  21. 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]
  22. Saraste J., Palade G. E., Farquhar M. G. Antibodies to rat pancreas Golgi subfractions: identification of a 58-kD cis-Golgi protein. J Cell Biol. 1987 Nov;105(5):2021–2029. doi: 10.1083/jcb.105.5.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schnabel E., Dekan G., Miettinen A., Farquhar M. G. Biogenesis of podocalyxin--the major glomerular sialoglycoprotein--in the newborn rat kidney. Eur J Cell Biol. 1989 Apr;48(2):313–326. [PubMed] [Google Scholar]
  24. Schnabel E., Mains R. E., Farquhar M. G. Proteolytic processing of pro-ACTH/endorphin begins in the Golgi complex of pituitary corticotropes and AtT-20 cells. Mol Endocrinol. 1989 Aug;3(8):1223–1235. doi: 10.1210/mend-3-8-1223. [DOI] [PubMed] [Google Scholar]
  25. Schweizer A., Fransen J. A., Bächi T., Ginsel L., Hauri H. P. Identification, by a monoclonal antibody, of a 53-kD protein associated with a tubulo-vesicular compartment at the cis-side of the Golgi apparatus. J Cell Biol. 1988 Nov;107(5):1643–1653. doi: 10.1083/jcb.107.5.1643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shaper N. L., Hollis G. F., Douglas J. G., Kirsch I. R., Shaper J. H. Characterization of the full length cDNA for murine beta-1,4-galactosyltransferase. Novel features at the 5'-end predict two translational start sites at two in-frame AUGs. J Biol Chem. 1988 Jul 25;263(21):10420–10428. [PubMed] [Google Scholar]
  27. Stern D. F., Sefton B. M. Coronavirus proteins: structure and function of the oligosaccharides of the avian infectious bronchitis virus glycoproteins. J Virol. 1982 Dec;44(3):804–812. doi: 10.1128/jvi.44.3.804-812.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tokuyasu K. T. Use of poly(vinylpyrrolidone) and poly(vinyl alcohol) for cryoultramicrotomy. Histochem J. 1989 Mar;21(3):163–171. doi: 10.1007/BF01007491. [DOI] [PubMed] [Google Scholar]
  29. Tooze J., Tooze S. A. Infection of AtT20 murine pituitary tumour cells by mouse hepatitis virus strain A59: virus budding is restricted to the Golgi region. Eur J Cell Biol. 1985 May;37:203–212. [PubMed] [Google Scholar]
  30. Tooze J., Tooze S., Warren G. Replication of coronavirus MHV-A59 in sac- cells: determination of the first site of budding of progeny virions. Eur J Cell Biol. 1984 Mar;33(2):281–293. [PubMed] [Google Scholar]
  31. Tooze S. A., Tooze J., Warren G. Site of addition of N-acetyl-galactosamine to the E1 glycoprotein of mouse hepatitis virus-A59. J Cell Biol. 1988 May;106(5):1475–1487. doi: 10.1083/jcb.106.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weinstein J., Lee E. U., McEntee K., Lai P. H., Paulson J. C. Primary structure of beta-galactoside alpha 2,6-sialyltransferase. Conversion of membrane-bound enzyme to soluble forms by cleavage of the NH2-terminal signal anchor. J Biol Chem. 1987 Dec 25;262(36):17735–17743. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES