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. 1994 Oct 17;13(20):4896–4907. doi: 10.1002/j.1460-2075.1994.tb06817.x

The functioning of the yeast Golgi apparatus requires an ER protein encoded by ANP1, a member of a new family of genes affecting the secretory pathway.

R E Chapman 1, S Munro 1
PMCID: PMC395430  PMID: 7957057

Abstract

Mnt1p is an alpha 1.2-mannosyltransferase which resides in an early compartment of the Saccharomyces cerevisiae Golgi apparatus. We have shown that the signal-anchor region is sufficient, and the transmembrane domain necessary, for its normal Golgi localization. This is similar to the transmembrane domain-mediated retention of mammalian glycosyltransferases, and distinct from the tail-mediated recycling retention of certain mammalian and yeast trans-Golgi proteins. To examine the mechanism involved in transmembrane domain-mediated retention, we have isolated six classes of mutants which fail to retain Mnt1p-reporter fusions in the early Golgi. These mutants all show additional phenotypes which are consistent with alterations in Golgi function. We have called the mutant classes 'gem', for Golgi enzyme maintenance. GEM3 is identical to the previously cloned gene ANP1, and homologous to VAN1 and MNN9. Together, these define a new class of proteins involved in the organization and functioning of the secretory pathway. Interestingly, Anp1p is localized to the endoplasmic reticulum (ER), implying that some function of the ER is required to maintain a functional Golgi apparatus.

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Selected References

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

  1. Ballou L., Hitzeman R. A., Lewis M. S., Ballou C. E. Vanadate-resistant yeast mutants are defective in protein glycosylation. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3209–3212. doi: 10.1073/pnas.88.8.3209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bos K., Wraight C., Stanley K. K. TGN38 is maintained in the trans-Golgi network by a tyrosine-containing motif in the cytoplasmic domain. EMBO J. 1993 May;12(5):2219–2228. doi: 10.1002/j.1460-2075.1993.tb05870.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bretscher M. S., Munro S. Cholesterol and the Golgi apparatus. Science. 1993 Sep 3;261(5126):1280–1281. doi: 10.1126/science.8362242. [DOI] [PubMed] [Google Scholar]
  4. Bryant N. J., Boyd A. Immunoisolation of Kex2p-containing organelles from yeast demonstrates colocalisation of three processing proteinases to a single Golgi compartment. J Cell Sci. 1993 Nov;106(Pt 3):815–822. doi: 10.1242/jcs.106.3.815. [DOI] [PubMed] [Google Scholar]
  5. Burke J., Pettitt J. M., Humphris D., Gleeson P. A. Medial-Golgi retention of N-acetylglucosaminyltransferase I. Contribution from all domains of the enzyme. J Biol Chem. 1994 Apr 22;269(16):12049–12059. [PubMed] [Google Scholar]
  6. Bussey H., Sacks W., Galley D., Saville D. Yeast killer plasmid mutations affecting toxin secretion and activity and toxin immunity function. Mol Cell Biol. 1982 Apr;2(4):346–354. doi: 10.1128/mcb.2.4.346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Camirand A., Heysen A., Grondin B., Herscovics A. Glycoprotein biosynthesis in Saccharomyces cerevisiae. Isolation and characterization of the gene encoding a specific processing alpha-mannosidase. J Biol Chem. 1991 Aug 15;266(23):15120–15127. [PubMed] [Google Scholar]
  8. Chapman R. E., Munro S. Retrieval of TGN proteins from the cell surface requires endosomal acidification. EMBO J. 1994 May 15;13(10):2305–2312. doi: 10.1002/j.1460-2075.1994.tb06514.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cooper A., Bussey H. Yeast Kex1p is a Golgi-associated membrane protein: deletions in a cytoplasmic targeting domain result in mislocalization to the vacuolar membrane. J Cell Biol. 1992 Dec;119(6):1459–1468. doi: 10.1083/jcb.119.6.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cunningham K. W., Wickner W. T. Yeast KEX2 protease and mannosyltransferase I are localized to distinct compartments of the secretory pathway. Yeast. 1989 Jan-Feb;5(1):25–33. doi: 10.1002/yea.320050105. [DOI] [PubMed] [Google Scholar]
  11. Dahdal R. Y., Colley K. J. Specific sequences in the signal anchor of the beta-galactoside alpha-2,6-sialyltransferase are not essential for Golgi localization. Membrane flanking sequences may specify Golgi retention. J Biol Chem. 1993 Dec 15;268(35):26310–26319. [PubMed] [Google Scholar]
  12. Franzusoff A., Schekman R. Functional compartments of the yeast Golgi apparatus are defined by the sec7 mutation. EMBO J. 1989 Sep;8(9):2695–2702. doi: 10.1002/j.1460-2075.1989.tb08410.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gopal P. K., Ballou C. E. Regulation of the protein glycosylation pathway in yeast: structural control of N-linked oligosaccharide elongation. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8824–8828. doi: 10.1073/pnas.84.24.8824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Graham T. R., Emr S. D. Compartmental organization of Golgi-specific protein modification and vacuolar protein sorting events defined in a yeast sec18 (NSF) mutant. J Cell Biol. 1991 Jul;114(2):207–218. doi: 10.1083/jcb.114.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Graham T. R., Scott P. A., Emr S. D. Brefeldin A reversibly blocks early but not late protein transport steps in the yeast secretory pathway. EMBO J. 1993 Mar;12(3):869–877. doi: 10.1002/j.1460-2075.1993.tb05727.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hardwick K. G., Boothroyd J. C., Rudner A. D., Pelham H. R. Genes that allow yeast cells to grow in the absence of the HDEL receptor. EMBO J. 1992 Nov;11(11):4187–4195. doi: 10.1002/j.1460-2075.1992.tb05512.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hardwick K. G., Lewis M. J., Semenza J., Dean N., Pelham H. R. ERD1, a yeast gene required for the retention of luminal endoplasmic reticulum proteins, affects glycoprotein processing in the Golgi apparatus. EMBO J. 1990 Mar;9(3):623–630. doi: 10.1002/j.1460-2075.1990.tb08154.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hardwick K. G., Pelham H. R. SED5 encodes a 39-kD integral membrane protein required for vesicular transport between the ER and the Golgi complex. J Cell Biol. 1992 Nov;119(3):513–521. doi: 10.1083/jcb.119.3.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hill K., Boone C., Goebl M., Puccia R., Sdicu A. M., Bussey H. Yeast KRE2 defines a new gene family encoding probable secretory proteins, and is required for the correct N-glycosylation of proteins. Genetics. 1992 Feb;130(2):273–283. doi: 10.1093/genetics/130.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Horazdovsky B. F., Emr S. D. The VPS16 gene product associates with a sedimentable protein complex and is essential for vacuolar protein sorting in yeast. J Biol Chem. 1993 Mar 5;268(7):4953–4962. [PubMed] [Google Scholar]
  21. Huffaker T. C., Robbins P. W. Temperature-sensitive yeast mutants deficient in asparagine-linked glycosylation. J Biol Chem. 1982 Mar 25;257(6):3203–3210. [PubMed] [Google Scholar]
  22. Huffaker T. C., Robbins P. W. Yeast mutants deficient in protein glycosylation. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7466–7470. doi: 10.1073/pnas.80.24.7466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Humphrey J. S., Peters P. J., Yuan L. C., Bonifacino J. S. Localization of TGN38 to the trans-Golgi network: involvement of a cytoplasmic tyrosine-containing sequence. J Cell Biol. 1993 Mar;120(5):1123–1135. doi: 10.1083/jcb.120.5.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Häusler A., Ballou L., Ballou C. E., Robbins P. W. Yeast glycoprotein biosynthesis: MNT1 encodes an alpha-1,2-mannosyltransferase involved in O-glycosylation. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6846–6850. doi: 10.1073/pnas.89.15.6846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Häusler A., Robbins P. W. Glycosylation in Saccharomyces cerevisiae: cloning and characterization of an alpha-1,2-mannosyltransferase structural gene. Glycobiology. 1992 Feb;2(1):77–84. doi: 10.1093/glycob/2.1.77. [DOI] [PubMed] [Google Scholar]
  26. Johnson K. F., Kornfeld S. The cytoplasmic tail of the mannose 6-phosphate/insulin-like growth factor-II receptor has two signals for lysosomal enzyme sorting in the Golgi. J Cell Biol. 1992 Oct;119(2):249–257. doi: 10.1083/jcb.119.2.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kaneko Y., Toh-e A., Oshima Y. Identification of the genetic locus for the structural gene and a new regulatory gene for the synthesis of repressible alkaline phosphatase in Saccharomyces cerevisiae. Mol Cell Biol. 1982 Feb;2(2):127–137. doi: 10.1128/mcb.2.2.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kanik-Ennulat C., Neff N. Vanadate-resistant mutants of Saccharomyces cerevisiae show alterations in protein phosphorylation and growth control. Mol Cell Biol. 1990 Mar;10(3):898–909. doi: 10.1128/mcb.10.3.898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Klionsky D. J., Emr S. D. Membrane protein sorting: biosynthesis, transport and processing of yeast vacuolar alkaline phosphatase. EMBO J. 1989 Aug;8(8):2241–2250. doi: 10.1002/j.1460-2075.1989.tb08348.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lewis M. S., Ballou C. E. Separation and characterization of two alpha 1,2-mannosyltransferase activities from Saccharomyces cerevisiae. J Biol Chem. 1991 May 5;266(13):8255–8261. [PubMed] [Google Scholar]
  31. Machamer C. E., Mentone S. A., Rose J. K., Farquhar M. G. The E1 glycoprotein of an avian coronavirus is targeted to the cis Golgi complex. Proc Natl Acad Sci U S A. 1990 Sep;87(18):6944–6948. doi: 10.1073/pnas.87.18.6944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Machamer C. E. Targeting and retention of Golgi membrane proteins. Curr Opin Cell Biol. 1993 Aug;5(4):606–612. doi: 10.1016/0955-0674(93)90129-E. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. McGee T. P., Skinner H. B., Whitters E. A., Henry S. A., Bankaitis V. A. A phosphatidylinositol transfer protein controls the phosphatidylcholine content of yeast Golgi membranes. J Cell Biol. 1994 Feb;124(3):273–287. doi: 10.1083/jcb.124.3.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. McKnight G. L., Cardillo T. S., Sherman F. An extensive deletion causing overproduction of yeast iso-2-cytochrome c. Cell. 1981 Aug;25(2):409–419. doi: 10.1016/0092-8674(81)90059-3. [DOI] [PubMed] [Google Scholar]
  35. Melnick L., Sherman F. The gene clusters ARC and COR on chromosomes 5 and 10, respectively, of Saccharomyces cerevisiae share a common ancestry. J Mol Biol. 1993 Oct 5;233(3):372–388. doi: 10.1006/jmbi.1993.1518. [DOI] [PubMed] [Google Scholar]
  36. Munro S., Pelham H. R. A C-terminal signal prevents secretion of luminal ER proteins. Cell. 1987 Mar 13;48(5):899–907. doi: 10.1016/0092-8674(87)90086-9. [DOI] [PubMed] [Google Scholar]
  37. Munro S. Sequences within and adjacent to the transmembrane segment of alpha-2,6-sialyltransferase specify Golgi retention. EMBO J. 1991 Dec;10(12):3577–3588. doi: 10.1002/j.1460-2075.1991.tb04924.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Nakanishi-Shindo Y., Nakayama K., Tanaka A., Toda Y., Jigami Y. Structure of the N-linked oligosaccharides that show the complete loss of alpha-1,6-polymannose outer chain from och1, och1 mnn1, and och1 mnn1 alg3 mutants of Saccharomyces cerevisiae. J Biol Chem. 1993 Dec 15;268(35):26338–26345. [PubMed] [Google Scholar]
  39. Nakayama K., Nagasu T., Shimma Y., Kuromitsu J., Jigami Y. OCH1 encodes a novel membrane bound mannosyltransferase: outer chain elongation of asparagine-linked oligosaccharides. EMBO J. 1992 Jul;11(7):2511–2519. doi: 10.1002/j.1460-2075.1992.tb05316.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Nilsson T., Lucocq J. M., Mackay D., Warren G. The membrane spanning domain of beta-1,4-galactosyltransferase specifies trans Golgi localization. EMBO J. 1991 Dec;10(12):3567–3575. doi: 10.1002/j.1460-2075.1991.tb04923.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Nilsson T., Slusarewicz P., Hoe M. H., Warren G. Kin recognition. A model for the retention of Golgi enzymes. FEBS Lett. 1993 Sep 6;330(1):1–4. doi: 10.1016/0014-5793(93)80906-b. [DOI] [PubMed] [Google Scholar]
  42. Nothwehr S. F., Roberts C. J., Stevens T. H. Membrane protein retention in the yeast Golgi apparatus: dipeptidyl aminopeptidase A is retained by a cytoplasmic signal containing aromatic residues. J Cell Biol. 1993 Jun;121(6):1197–1209. doi: 10.1083/jcb.121.6.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Pelham H. R. Control of protein exit from the endoplasmic reticulum. Annu Rev Cell Biol. 1989;5:1–23. doi: 10.1146/annurev.cb.05.110189.000245. [DOI] [PubMed] [Google Scholar]
  44. Pelham H. R., Hardwick K. G., Lewis M. J. Sorting of soluble ER proteins in yeast. EMBO J. 1988 Jun;7(6):1757–1762. doi: 10.1002/j.1460-2075.1988.tb03005.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Pelham H. R., Munro S. Sorting of membrane proteins in the secretory pathway. Cell. 1993 Nov 19;75(4):603–605. doi: 10.1016/0092-8674(93)90479-A. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Pfeffer S. R., Rothman J. E. Biosynthetic protein transport and sorting by the endoplasmic reticulum and Golgi. Annu Rev Biochem. 1987;56:829–852. doi: 10.1146/annurev.bi.56.070187.004145. [DOI] [PubMed] [Google Scholar]
  47. Raschke W. C., Kern K. A., Antalis C., Ballou C. E. Genetic control of yeast mannan structure. Isolation and characterization of mannan mutants. J Biol Chem. 1973 Jul 10;248(13):4660–4666. [PubMed] [Google Scholar]
  48. Roberts C. J., Raymond C. K., Yamashiro C. T., Stevens T. H. Methods for studying the yeast vacuole. Methods Enzymol. 1991;194:644–661. doi: 10.1016/0076-6879(91)94047-g. [DOI] [PubMed] [Google Scholar]
  49. Rudolph H. K., Antebi A., Fink G. R., Buckley C. M., Dorman T. E., LeVitre J., Davidow L. S., Mao J. I., Moir D. T. The yeast secretory pathway is perturbed by mutations in PMR1, a member of a Ca2+ ATPase family. Cell. 1989 Jul 14;58(1):133–145. doi: 10.1016/0092-8674(89)90410-8. [DOI] [PubMed] [Google Scholar]
  50. Sweet D. J., Pelham H. R. The TIP1 gene of Saccharomyces cerevisiae encodes an 80 kDa cytoplasmic protein that interacts with the cytoplasmic domain of Sec20p. EMBO J. 1993 Jul;12(7):2831–2840. doi: 10.1002/j.1460-2075.1993.tb05944.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Vogel J. P., Lee J. N., Kirsch D. R., Rose M. D., Sztul E. S. Brefeldin A causes a defect in secretion in Saccharomyces cerevisiae. J Biol Chem. 1993 Feb 15;268(5):3040–3043. [PubMed] [Google Scholar]
  52. Weisz O. A., Swift A. M., Machamer C. E. Oligomerization of a membrane protein correlates with its retention in the Golgi complex. J Cell Biol. 1993 Sep;122(6):1185–1196. doi: 10.1083/jcb.122.6.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wilcox C. A., Redding K., Wright R., Fuller R. S. Mutation of a tyrosine localization signal in the cytosolic tail of yeast Kex2 protease disrupts Golgi retention and results in default transport to the vacuole. Mol Biol Cell. 1992 Dec;3(12):1353–1371. doi: 10.1091/mbc.3.12.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wilsbach K., Payne G. S. Vps1p, a member of the dynamin GTPase family, is necessary for Golgi membrane protein retention in Saccharomyces cerevisiae. EMBO J. 1993 Aug;12(8):3049–3059. doi: 10.1002/j.1460-2075.1993.tb05974.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Wong S. H., Hong W. The SXYQRL sequence in the cytoplasmic domain of TGN38 plays a major role in trans-Golgi network localization. J Biol Chem. 1993 Oct 25;268(30):22853–22862. [PubMed] [Google Scholar]
  56. Yip C. L., Welch S. K., Klebl F., Gilbert T., Seidel P., Grant F. J., O'Hara P. J., MacKay V. L. Cloning and analysis of the Saccharomyces cerevisiae MNN9 and MNN1 genes required for complex glycosylation of secreted proteins. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2723–2727. doi: 10.1073/pnas.91.7.2723. [DOI] [PMC free article] [PubMed] [Google Scholar]

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