Skip to main content
NCBI 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
. 1995 Apr 11;92(8):3229–3233. doi: 10.1073/pnas.92.8.3229

Non-clathrin-coat protein alpha is a conserved subunit of coatomer and in Saccharomyces cerevisiae is essential for growth.

B Gerich 1, L Orci 1, H Tschochner 1, F Lottspeich 1, M Ravazzola 1, M Amherdt 1, F Wieland 1, C Harter 1
PMCID: PMC42139  PMID: 7724544

Abstract

To complete the molecular characterization of coatomer, the preformed cytosolic complex that is involved in the formation of biosynthetic transport vesicles, we have cloned and characterized the gene for non-clathrin-coat protein alpha (alpha-COP) from Saccharomyces cerevisiae. The derived protein, molecular weight of 135,500, contains four WD-40 repeated motifs (Trp/Asp-containing motifs of approximately 40 amino acids). Disruption of the yeast alpha-COP gene is lethal. Comparison of the DNA-derived primary structure with peptides from bovine alpha-COP shows a striking homology. alpha-COP is localized to coated transport vesicles and coated buds of Golgi membranes derived from CHO cells.

Full text

PDF
3229

Images in this article

3231Fig. 3
on p.3231
3232Fig. 4
on p.3232
3232Fig. 5
on p.3232

Selected References

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

  1. Barlowe C., Orci L., Yeung T., Hosobuchi M., Hamamoto S., Salama N., Rexach M. F., Ravazzola M., Amherdt M., Schekman R. COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum. Cell. 1994 Jun 17;77(6):895–907. doi: 10.1016/0092-8674(94)90138-4. [DOI] [PubMed] [Google Scholar]
  2. Cosson P., Letourneur F. Coatomer interaction with di-lysine endoplasmic reticulum retention motifs. Science. 1994 Mar 18;263(5153):1629–1631. doi: 10.1126/science.8128252. [DOI] [PubMed] [Google Scholar]
  3. Donaldson J. G., Cassel D., Kahn R. A., Klausner R. D. ADP-ribosylation factor, a small GTP-binding protein, is required for binding of the coatomer protein beta-COP to Golgi membranes. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6408–6412. doi: 10.1073/pnas.89.14.6408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Duden R., Griffiths G., Frank R., Argos P., Kreis T. E. Beta-COP, a 110 kd protein associated with non-clathrin-coated vesicles and the Golgi complex, shows homology to beta-adaptin. Cell. 1991 Feb 8;64(3):649–665. doi: 10.1016/0092-8674(91)90248-w. [DOI] [PubMed] [Google Scholar]
  5. Duden R., Hosobuchi M., Hamamoto S., Winey M., Byers B., Schekman R. Yeast beta- and beta'-coat proteins (COP). Two coatomer subunits essential for endoplasmic reticulum-to-Golgi protein traffic. J Biol Chem. 1994 Sep 30;269(39):24486–24495. [PubMed] [Google Scholar]
  6. Gietz D., St Jean A., Woods R. A., Schiestl R. H. Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res. 1992 Mar 25;20(6):1425–1425. doi: 10.1093/nar/20.6.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Guo Q., Vasile E., Krieger M. Disruptions in Golgi structure and membrane traffic in a conditional lethal mammalian cell mutant are corrected by epsilon-COP. J Cell Biol. 1994 Jun;125(6):1213–1224. doi: 10.1083/jcb.125.6.1213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hara-Kuge S., Kuge O., Orci L., Amherdt M., Ravazzola M., Wieland F. T., Rothman J. E. En bloc incorporation of coatomer subunits during the assembly of COP-coated vesicles. J Cell Biol. 1994 Mar;124(6):883–892. doi: 10.1083/jcb.124.6.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Harrison-Lavoie K. J., Lewis V. A., Hynes G. M., Collison K. S., Nutland E., Willison K. R. A 102 kDa subunit of a Golgi-associated particle has homology to beta subunits of trimeric G proteins. EMBO J. 1993 Jul;12(7):2847–2853. doi: 10.1002/j.1460-2075.1993.tb05946.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harter C., Draken E., Lottspeich F., Wieland F. T. Yeast coatomer contains a subunit homologous to mammalian beta'-COP. FEBS Lett. 1993 Oct 11;332(1-2):71–73. doi: 10.1016/0014-5793(93)80487-f. [DOI] [PubMed] [Google Scholar]
  11. Hosobuchi M., Kreis T., Schekman R. SEC21 is a gene required for ER to Golgi protein transport that encodes a subunit of a yeast coatomer. Nature. 1992 Dec 10;360(6404):603–605. doi: 10.1038/360603a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Kuge O., Hara-Kuge S., Orci L., Ravazzola M., Amherdt M., Tanigawa G., Wieland F. T., Rothman J. E. zeta-COP, a subunit of coatomer, is required for COP-coated vesicle assembly. J Cell Biol. 1993 Dec;123(6 Pt 2):1727–1734. doi: 10.1083/jcb.123.6.1727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Neer E. J., Schmidt C. J., Nambudripad R., Smith T. F. The ancient regulatory-protein family of WD-repeat proteins. Nature. 1994 Sep 22;371(6495):297–300. doi: 10.1038/371297a0. [DOI] [PubMed] [Google Scholar]
  15. Nilsson T., Jackson M., Peterson P. A. Short cytoplasmic sequences serve as retention signals for transmembrane proteins in the endoplasmic reticulum. Cell. 1989 Aug 25;58(4):707–718. doi: 10.1016/0092-8674(89)90105-0. [DOI] [PubMed] [Google Scholar]
  16. Orci L., Glick B. S., Rothman J. E. A new type of coated vesicular carrier that appears not to contain clathrin: its possible role in protein transport within the Golgi stack. Cell. 1986 Jul 18;46(2):171–184. doi: 10.1016/0092-8674(86)90734-8. [DOI] [PubMed] [Google Scholar]
  17. Orci L., Malhotra V., Amherdt M., Serafini T., Rothman J. E. Dissection of a single round of vesicular transport: sequential intermediates for intercisternal movement in the Golgi stack. Cell. 1989 Feb 10;56(3):357–368. doi: 10.1016/0092-8674(89)90239-0. [DOI] [PubMed] [Google Scholar]
  18. Orci L., Palmer D. J., Ravazzola M., Perrelet A., Amherdt M., Rothman J. E. Budding from Golgi membranes requires the coatomer complex of non-clathrin coat proteins. Nature. 1993 Apr 15;362(6421):648–652. doi: 10.1038/362648a0. [DOI] [PubMed] [Google Scholar]
  19. Orci L., Ravazzola M., Meda P., Holcomb C., Moore H. P., Hicke L., Schekman R. Mammalian Sec23p homologue is restricted to the endoplasmic reticulum transitional cytoplasm. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8611–8615. doi: 10.1073/pnas.88.19.8611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Orcl L., Palmer D. J., Amherdt M., Rothman J. E. Coated vesicle assembly in the Golgi requires only coatomer and ARF proteins from the cytosol. Nature. 1993 Aug 19;364(6439):732–734. doi: 10.1038/364732a0. [DOI] [PubMed] [Google Scholar]
  21. Ostermann J., Orci L., Tani K., Amherdt M., Ravazzola M., Elazar Z., Rothman J. E. Stepwise assembly of functionally active transport vesicles. Cell. 1993 Dec 3;75(5):1015–1025. doi: 10.1016/0092-8674(93)90545-2. [DOI] [PubMed] [Google Scholar]
  22. Palmer D. J., Helms J. B., Beckers C. J., Orci L., Rothman J. E. Binding of coatomer to Golgi membranes requires ADP-ribosylation factor. J Biol Chem. 1993 Jun 5;268(16):12083–12089. [PubMed] [Google Scholar]
  23. Pepperkok R., Scheel J., Horstmann H., Hauri H. P., Griffiths G., Kreis T. E. Beta-COP is essential for biosynthetic membrane transport from the endoplasmic reticulum to the Golgi complex in vivo. Cell. 1993 Jul 16;74(1):71–82. doi: 10.1016/0092-8674(93)90295-2. [DOI] [PubMed] [Google Scholar]
  24. Posnett D. N., Tam J. P. Multiple antigenic peptide method for producing antipeptide site-specific antibodies. Methods Enzymol. 1989;178:739–746. doi: 10.1016/0076-6879(89)78048-4. [DOI] [PubMed] [Google Scholar]
  25. Pryer N. K., Salama N. R., Schekman R., Kaiser C. A. Cytosolic Sec13p complex is required for vesicle formation from the endoplasmic reticulum in vitro. J Cell Biol. 1993 Feb;120(4):865–875. doi: 10.1083/jcb.120.4.865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pryer N. K., Wuestehube L. J., Schekman R. Vesicle-mediated protein sorting. Annu Rev Biochem. 1992;61:471–516. doi: 10.1146/annurev.bi.61.070192.002351. [DOI] [PubMed] [Google Scholar]
  27. Rexach M. F., Latterich M., Schekman R. W. Characteristics of endoplasmic reticulum-derived transport vesicles. J Cell Biol. 1994 Sep;126(5):1133–1148. doi: 10.1083/jcb.126.5.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Roth J., Bendayan M., Orci L. Ultrastructural localization of intracellular antigens by the use of protein A-gold complex. J Histochem Cytochem. 1978 Dec;26(12):1074–1081. doi: 10.1177/26.12.366014. [DOI] [PubMed] [Google Scholar]
  29. Rothman J. E., Orci L. Molecular dissection of the secretory pathway. Nature. 1992 Jan 30;355(6359):409–415. doi: 10.1038/355409a0. [DOI] [PubMed] [Google Scholar]
  30. Serafini T., Orci L., Amherdt M., Brunner M., Kahn R. A., Rothman J. E. ADP-ribosylation factor is a subunit of the coat of Golgi-derived COP-coated vesicles: a novel role for a GTP-binding protein. Cell. 1991 Oct 18;67(2):239–253. doi: 10.1016/0092-8674(91)90176-y. [DOI] [PubMed] [Google Scholar]
  31. Serafini T., Stenbeck G., Brecht A., Lottspeich F., Orci L., Rothman J. E., Wieland F. T. A coat subunit of Golgi-derived non-clathrin-coated vesicles with homology to the clathrin-coated vesicle coat protein beta-adaptin. Nature. 1991 Jan 17;349(6306):215–220. doi: 10.1038/349215a0. [DOI] [PubMed] [Google Scholar]
  32. Stenbeck G., Harter C., Brecht A., Herrmann D., Lottspeich F., Orci L., Wieland F. T. beta'-COP, a novel subunit of coatomer. EMBO J. 1993 Jul;12(7):2841–2845. doi: 10.1002/j.1460-2075.1993.tb05945.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stenbeck G., Schreiner R., Herrmann D., Auerbach S., Lottspeich F., Rothman J. E., Wieland F. T. Gamma-COP, a coat subunit of non-clathrin-coated vesicles with homology to Sec21p. FEBS Lett. 1992 Dec 14;314(2):195–198. doi: 10.1016/0014-5793(92)80973-k. [DOI] [PubMed] [Google Scholar]
  34. Tokuyasu K. T. Immunochemistry on ultrathin frozen sections. Histochem J. 1980 Jul;12(4):381–403. doi: 10.1007/BF01011956. [DOI] [PubMed] [Google Scholar]
  35. Vinh D. B., Drubin D. G. A yeast TCP-1-like protein is required for actin function in vivo. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9116–9120. doi: 10.1073/pnas.91.19.9116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Waters M. G., Serafini T., Rothman J. E. 'Coatomer': a cytosolic protein complex containing subunits of non-clathrin-coated Golgi transport vesicles. Nature. 1991 Jan 17;349(6306):248–251. doi: 10.1038/349248a0. [DOI] [PubMed] [Google Scholar]
  37. van der Voorn L., Ploegh H. L. The WD-40 repeat. FEBS Lett. 1992 Jul 28;307(2):131–134. doi: 10.1016/0014-5793(92)80751-2. [DOI] [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