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. 1993 Sep;5(9):1113–1124. doi: 10.1105/tpc.5.9.1113

Tonoplast and Soluble Vacuolar Proteins Are Targeted by Different Mechanisms.

L Gomez 1, MJ Chrispeels 1
PMCID: PMC160345  PMID: 12271099

Abstract

The delivery of proteins to the vacuole and its limiting membrane (the tonoplast) by the secretory system is thought to be a dissociative process in which vesicles bud from one compartment and fuse with another. We studied the transport kinetics of phytohemagglutinin (PHA) and tonoplast intrinsic protein (TIP) in mesophyll protoplasts obtained from transgenic tobacco plants transformed with genes encoding these two proteins. In pulse-chase experiments, arrival of PHA in the vacuole was found to be slower (completed 24 hr after synthesis) than the arrival of TIP in the tonoplast (completed 6 hr after synthesis). Brefeldin A and monensin block protein transport by interfering in specific vesicle transport steps. Brefeldin A prevents anterograde vesicle transport between the endoplasmic reticulum and the Golgi, whereas monensin inhibits correct sorting in the trans-Golgi network by disrupting the proton gradient across the membrane. Both inhibitors blocked the transport of PHA to the vacuole and altered the rate at which its complex glycan is processed by Golgi enzymes. Neither drug stopped the arrival of TIP in the tonoplast, suggesting that the flow of vesicles continues in the presence of these inhibitors. We suggest that soluble proteins like PHA and membrane proteins like TIP reach their vacuolar destinations by different paths.

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Selected References

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  1. Anderson R. G., Pathak R. K. Vesicles and cisternae in the trans Golgi apparatus of human fibroblasts are acidic compartments. Cell. 1985 Mar;40(3):635–643. doi: 10.1016/0092-8674(85)90212-0. [DOI] [PubMed] [Google Scholar]
  2. Bednarek S. Y., Raikhel N. V. Intracellular trafficking of secretory proteins. Plant Mol Biol. 1992 Oct;20(1):133–150. doi: 10.1007/BF00029156. [DOI] [PubMed] [Google Scholar]
  3. Bowles D. J., Marcus S. E., Pappin D. J., Findlay J. B., Eliopoulos E., Maycox P. R., Burgess J. Posttranslational processing of concanavalin A precursors in jackbean cotyledons. J Cell Biol. 1986 Apr;102(4):1284–1297. doi: 10.1083/jcb.102.4.1284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chawla D., Hughes R. C. Effects of brefeldin A on oligosaccharide processing. Evidence for decreased branching of complex-type glycans and increased formation of hybrid-type glycans. Biochem J. 1991 Oct 1;279(Pt 1):159–165. doi: 10.1042/bj2790159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chrispeels M. J., Raikhel N. V. Short peptide domains target proteins to plant vacuoles. Cell. 1992 Feb 21;68(4):613–616. doi: 10.1016/0092-8674(92)90134-x. [DOI] [PubMed] [Google Scholar]
  6. Collins P. L., Mottet G. Oligomerization and post-translational processing of glycoprotein G of human respiratory syncytial virus: altered O-glycosylation in the presence of brefeldin A. J Gen Virol. 1992 Apr;73(Pt 4):849–863. doi: 10.1099/0022-1317-73-4-849. [DOI] [PubMed] [Google Scholar]
  7. Denecke J., Botterman J., Deblaere R. Protein secretion in plant cells can occur via a default pathway. Plant Cell. 1990 Jan;2(1):51–59. doi: 10.1105/tpc.2.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Donaldson J. G., Finazzi D., Klausner R. D. Brefeldin A inhibits Golgi membrane-catalysed exchange of guanine nucleotide onto ARF protein. Nature. 1992 Nov 26;360(6402):350–352. doi: 10.1038/360350a0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Greenwood J. S., Chrispeels M. J. Correct targeting of the bean storage protein phaseolin in the seeds of transformed tobacco. Plant Physiol. 1985 Sep;79(1):65–71. doi: 10.1104/pp.79.1.65. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Harley S. M., Beevers L. Coated Vesicles Are Involved in the Transport of Storage Proteins during Seed Development in Pisum sativum L. Plant Physiol. 1989 Oct;91(2):674–678. doi: 10.1104/pp.91.2.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Helms J. B., Rothman J. E. Inhibition by brefeldin A of a Golgi membrane enzyme that catalyses exchange of guanine nucleotide bound to ARF. Nature. 1992 Nov 26;360(6402):352–354. doi: 10.1038/360352a0. [DOI] [PubMed] [Google Scholar]
  13. Holwerda B. C., Galvin N. J., Baranski T. J., Rogers J. C. In Vitro Processing of Aleurain, a Barley Vacuolar Thiol Protease. Plant Cell. 1990 Nov;2(11):1091–1106. doi: 10.1105/tpc.2.11.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Holwerda B. C., Padgett H. S., Rogers J. C. Proaleurain vacuolar targeting is mediated by short contiguous peptide interactions. Plant Cell. 1992 Mar;4(3):307–318. doi: 10.1105/tpc.4.3.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hunt D. C., Chrispeels M. J. The signal Peptide of a vacuolar protein is necessary and sufficient for the efficient secretion of a cytosolic protein. Plant Physiol. 1991 May;96(1):18–25. doi: 10.1104/pp.96.1.18. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Höfgen R., Willmitzer L. Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res. 1988 Oct 25;16(20):9877–9877. doi: 10.1093/nar/16.20.9877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Höfte H., Chrispeels M. J. Protein sorting to the vacuolar membrane. Plant Cell. 1992 Aug;4(8):995–1004. doi: 10.1105/tpc.4.8.995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Höfte H., Hubbard L., Reizer J., Ludevid D., Herman E. M., Chrispeels M. J. Vegetative and Seed-Specific Forms of Tonoplast Intrinsic Protein in the Vacuolar Membrane of Arabidopsis thaliana. Plant Physiol. 1992 Jun;99(2):561–570. doi: 10.1104/pp.99.2.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jones A. M., Herman E. M. KDEL-Containing Auxin-Binding Protein Is Secreted to the Plasma Membrane and Cell Wall. Plant Physiol. 1993 Feb;101(2):595–606. doi: 10.1104/pp.101.2.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Klausner R. D., Donaldson J. G., Lippincott-Schwartz J. Brefeldin A: insights into the control of membrane traffic and organelle structure. J Cell Biol. 1992 Mar;116(5):1071–1080. doi: 10.1083/jcb.116.5.1071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Kornfeld S., Mellman I. The biogenesis of lysosomes. Annu Rev Cell Biol. 1989;5:483–525. doi: 10.1146/annurev.cb.05.110189.002411. [DOI] [PubMed] [Google Scholar]
  23. Ktistakis N. T., Roth M. G., Bloom G. S. PtK1 cells contain a nondiffusible, dominant factor that makes the Golgi apparatus resistant to brefeldin A. J Cell Biol. 1991 Jun;113(5):1009–1023. doi: 10.1083/jcb.113.5.1009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lainé A. C., Gomord V., Faye L. Xylose-specific antibodies as markers of subcompartmentation of terminal glycosylation in the Golgi apparatus of sycamore cells. FEBS Lett. 1991 Dec 16;295(1-3):179–184. doi: 10.1016/0014-5793(91)81413-3. [DOI] [PubMed] [Google Scholar]
  25. Lippincott-Schwartz J., Donaldson J. G., Schweizer A., Berger E. G., Hauri H. P., Yuan L. C., Klausner R. D. Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway. Cell. 1990 Mar 9;60(5):821–836. doi: 10.1016/0092-8674(90)90096-w. [DOI] [PubMed] [Google Scholar]
  26. Maurel C., Reizer J., Schroeder J. I., Chrispeels M. J. The vacuolar membrane protein gamma-TIP creates water specific channels in Xenopus oocytes. EMBO J. 1993 Jun;12(6):2241–2247. doi: 10.1002/j.1460-2075.1993.tb05877.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Misumi Y., Misumi Y., Miki K., Takatsuki A., Tamura G., Ikehara Y. Novel blockade by brefeldin A of intracellular transport of secretory proteins in cultured rat hepatocytes. J Biol Chem. 1986 Aug 25;261(24):11398–11403. [PubMed] [Google Scholar]
  28. Mollenhauer H. H., Morré D. J., Rowe L. D. Alteration of intracellular traffic by monensin; mechanism, specificity and relationship to toxicity. Biochim Biophys Acta. 1990 May 7;1031(2):225–246. doi: 10.1016/0304-4157(90)90008-Z. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Moore P. J., Swords K. M., Lynch M. A., Staehelin L. A. Spatial organization of the assembly pathways of glycoproteins and complex polysaccharides in the Golgi apparatus of plants. J Cell Biol. 1991 Feb;112(4):589–602. doi: 10.1083/jcb.112.4.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pelham H. R. Multiple targets for brefeldin A. Cell. 1991 Nov 1;67(3):449–451. doi: 10.1016/0092-8674(91)90517-3. [DOI] [PubMed] [Google Scholar]
  31. Raymond C. K., Howald-Stevenson I., Vater C. A., Stevens T. H. Morphological classification of the yeast vacuolar protein sorting mutants: evidence for a prevacuolar compartment in class E vps mutants. Mol Biol Cell. 1992 Dec;3(12):1389–1402. doi: 10.1091/mbc.3.12.1389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Roberts C. J., Nothwehr S. F., Stevens T. H. Membrane protein sorting in the yeast secretory pathway: evidence that the vacuole may be the default compartment. J Cell Biol. 1992 Oct;119(1):69–83. doi: 10.1083/jcb.119.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Sandvig K., Prydz K., Hansen S. H., van Deurs B. Ricin transport in brefeldin A-treated cells: correlation between Golgi structure and toxic effect. J Cell Biol. 1991 Nov;115(4):971–981. doi: 10.1083/jcb.115.4.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schu P. V., Takegawa K., Fry M. J., Stack J. H., Waterfield M. D., Emr S. D. Phosphatidylinositol 3-kinase encoded by yeast VPS34 gene essential for protein sorting. Science. 1993 Apr 2;260(5104):88–91. doi: 10.1126/science.8385367. [DOI] [PubMed] [Google Scholar]
  36. Seeger M., Payne G. S. Selective and immediate effects of clathrin heavy chain mutations on Golgi membrane protein retention in Saccharomyces cerevisiae. J Cell Biol. 1992 Aug;118(3):531–540. doi: 10.1083/jcb.118.3.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shah N., Klausner R. D. Brefeldin A reversibly inhibits secretion in Saccharomyces cerevisiae. J Biol Chem. 1993 Mar 15;268(8):5345–5348. [PubMed] [Google Scholar]
  38. Sonnewald U., Brauer M., von Schaewen A., Stitt M., Willmitzer L. Transgenic tobacco plants expressing yeast-derived invertase in either the cytosol, vacuole or apoplast: a powerful tool for studying sucrose metabolism and sink/source interactions. Plant J. 1991 Jul;1(1):95–106. doi: 10.1111/j.1365-313x.1991.00095.x. [DOI] [PubMed] [Google Scholar]
  39. Tague B. W., Dickinson C. D., Chrispeels M. J. A short domain of the plant vacuolar protein phytohemagglutinin targets invertase to the yeast vacuole. Plant Cell. 1990 Jun;2(6):533–546. doi: 10.1105/tpc.2.6.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Vitale A., Ceriotti A., Bollini R., Chrispeels M. J. Biosynthesis and processing of phytohemagglutinin in developing bean cotyledons. Eur J Biochem. 1984 May 15;141(1):97–104. doi: 10.1111/j.1432-1033.1984.tb08162.x. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. 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]
  43. Wong D. H., Brodsky F. M. 100-kD proteins of Golgi- and trans-Golgi network-associated coated vesicles have related but distinct membrane binding properties. J Cell Biol. 1992 Jun;117(6):1171–1179. doi: 10.1083/jcb.117.6.1171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zhang G. F., Driouich A., Staehelin L. A. Effect of monensin on plant Golgi: re-examination of the monensin-induced changes in cisternal architecture and functional activities of the Golgi apparatus of sycamore suspension-cultured cells. J Cell Sci. 1993 Mar;104(Pt 3):819–831. doi: 10.1242/jcs.104.3.819. [DOI] [PubMed] [Google Scholar]
  45. Zhang G. F., Staehelin L. A. Functional compartmentation of the Golgi apparatus of plant cells : immunocytochemical analysis of high-pressure frozen- and freeze-substituted sycamore maple suspension culture cells. Plant Physiol. 1992 Jul;99(3):1070–1083. doi: 10.1104/pp.99.3.1070. [DOI] [PMC free article] [PubMed] [Google Scholar]

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