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Plant Cell. Nov 1999; 11(11): 2233–2248.
PMCID: PMC144130

Saturation of the endoplasmic reticulum retention machinery reveals anterograde bulk flow

Abstract

We have studied the possible mechanisms of endoplasmic reticulum (ER) export and retention by using natural residents of the plant ER. Under normal physiological conditions, calreticulin and the lumenal binding protein (BiP) are efficiently retained in the ER. When the ER retention signal is removed, truncated calreticulin is much more rapidly secreted than truncated BiP. Calreticulin carries two glycans of the typical ER high-mannose form. Both glycans are competent for Golgi-based modifications, as determined from treatment with brefeldin A or based on the deletion of the ER retention motif. In contrast to BiP, calreticulin accumulation is strongly dependent on its retention signal, thereby allowing us to test whether saturation of the retention mechanism is possible. Overexpression of calreticulin led to 100-fold higher levels in dilated globular ER cisternae as well as dilated nuclear envelopes and partial secretion of both BiP and calreticulin. This result shows that both molecules are competent for ER export and supports the concept that proteins are secreted by default. This result also supports previous data suggesting that truncated BiP devoid of its retention motif can be retained in the ER by association with calreticulin. Moreover, even under these saturating conditions, cellular calreticulin did not carry significant amounts of complex glycans, in contrast to secreted calreticulin. This result shows that calreticulin is rapidly secreted once complex glycans have been synthesized in the medial/trans Golgi apparatus and that the modified protein does not appear to recycle back to the ER.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Balch WE, McCaffery JM, Plutner H, Farquhar MG. Vesicular stomatitis virus glycoprotein is sorted and concentrated during export from the endoplasmic reticulum. Cell. 1994 Mar 11;76(5):841–852. [PubMed]
  • Barlowe C, Orci L, Yeung T, Hosobuchi M, Hamamoto S, Salama N, Rexach MF, 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. [PubMed]
  • Boevink P, Oparka K, Santa Cruz S, Martin B, Betteridge A, Hawes C. Stacks on tracks: the plant Golgi apparatus traffics on an actin/ER network. Plant J. 1998 Aug;15(3):441–447. [PubMed]
  • Casadaban MJ, Cohen SN. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. [PubMed]
  • Ceriotti A, Colman A. Binding to membrane proteins within the endoplasmic reticulum cannot explain the retention of the glucose-regulated protein GRP78 in Xenopus oocytes. EMBO J. 1988 Mar;7(3):633–638. [PMC free article] [PubMed]
  • Connolly CN, Futter CE, Gibson A, Hopkins CR, Cutler DF. Transport into and out of the Golgi complex studied by transfecting cells with cDNAs encoding horseradish peroxidase. J Cell Biol. 1994 Nov;127(3):641–652. [PMC free article] [PubMed]
  • Cosson P, Letourneur F. Coatomer interaction with di-lysine endoplasmic reticulum retention motifs. Science. 1994 Mar 18;263(5153):1629–1631. [PubMed]
  • Crofts AJ, Leborgne-Castel N, Pesca M, Vitale A, Denecke J. BiP and calreticulin form an abundant complex that is independent of endoplasmic reticulum stress . Plant Cell. 1998 May;10(5):813–824. [PMC free article] [PubMed]
  • Dean N, Pelham HR. Recycling of proteins from the Golgi compartment to the ER in yeast. J Cell Biol. 1990 Aug;111(2):369–377. [PMC free article] [PubMed]
  • Deblaere R, Bytebier B, De Greve H, Deboeck F, Schell J, Van Montagu M, Leemans J. Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants. Nucleic Acids Res. 1985 Jul 11;13(13):4777–4788. [PMC free article] [PubMed]
  • Denecke J, Vitale A. The use of protoplasts to study protein synthesis and transport by the plant endomembrane system. Methods Cell Biol. 1995;50:335–348. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Denecke J, Goldman MH, Demolder J, Seurinck J, Botterman J. The tobacco luminal binding protein is encoded by a multigene family. Plant Cell. 1991 Sep;3(9):1025–1035. [PMC free article] [PubMed]
  • Denecke J, De Rycke R, Botterman J. Plant and mammalian sorting signals for protein retention in the endoplasmic reticulum contain a conserved epitope. EMBO J. 1992 Jun;11(6):2345–2355. [PMC free article] [PubMed]
  • Denecke J, Carlsson LE, Vidal S, Höglund AS, Ek B, van Zeijl MJ, Sinjorgo KM, Palva ET. The tobacco homolog of mammalian calreticulin is present in protein complexes in vivo. Plant Cell. 1995 Apr;7(4):391–406. [PMC free article] [PubMed]
  • Dorner AJ, Wasley LC, Raney P, Haugejorden S, Green M, Kaufman RJ. The stress response in Chinese hamster ovary cells. Regulation of ERp72 and protein disulfide isomerase expression and secretion. J Biol Chem. 1990 Dec 15;265(35):22029–22034. [PubMed]
  • Gomord V, Denmat LA, Fitchette-Lainé AC, Satiat-Jeunemaitre B, Hawes C, Faye L. The C-terminal HDEL sequence is sufficient for retention of secretory proteins in the endoplasmic reticulum (ER) but promotes vacuolar targeting of proteins that escape the ER. Plant J. 1997 Feb;11(2):313–325. [PubMed]
  • Hauri HP, Schweizer A. The endoplasmic reticulum-Golgi intermediate compartment. Curr Opin Cell Biol. 1992 Aug;4(4):600–608. [PubMed]
  • Hohl I, Robinson DG, Chrispeels MJ, Hinz G. Transport of storage proteins to the vacuole is mediated by vesicles without a clathrin coat. J Cell Sci. 1996 Oct;109(Pt 10):2539–2550. [PubMed]
  • Hong W. Protein transport from the endoplasmic reticulum to the Golgi apparatus. J Cell Sci. 1998 Oct;111(Pt 19):2831–2839. [PubMed]
  • Hunt DC, Chrispeels MJ. 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. [PMC free article] [PubMed]
  • Johannes L, Goud B. Surfing on a retrograde wave: how does Shiga toxin reach the endoplasmic reticulum? Trends Cell Biol. 1998 Apr;8(4):158–162. [PubMed]
  • Kornfeld R, Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. [PubMed]
  • Leborgne-Castel N, Jelitto-Van Dooren EP, Crofts AJ, Denecke J. Overexpression of BiP in tobacco alleviates endoplasmic reticulum stress. Plant Cell. 1999 Mar;11(3):459–470. [PMC free article] [PubMed]
  • Lee HI, Gal S, Newman TC, Raikhel NV. The Arabidopsis endoplasmic reticulum retention receptor functions in yeast. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11433–11437. [PMC free article] [PubMed]
  • Lewis MJ, Pelham HR. Ligand-induced redistribution of a human KDEL receptor from the Golgi complex to the endoplasmic reticulum. Cell. 1992 Jan 24;68(2):353–364. [PubMed]
  • Lippincott-Schwartz J, Donaldson JG, Schweizer A, Berger EG, Hauri HP, Yuan LC, Klausner RD. 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. [PubMed]
  • Lord JM, Roberts LM. Retrograde transport: going against the flow. Curr Biol. 1998 Jan 15;8(2):R56–R58. [PubMed]
  • Majoul I, Sohn K, Wieland FT, Pepperkok R, Pizza M, Hillemann J, Söling HD. KDEL receptor (Erd2p)-mediated retrograde transport of the cholera toxin A subunit from the Golgi involves COPI, p23, and the COOH terminus of Erd2p. J Cell Biol. 1998 Nov 2;143(3):601–612. [PMC free article] [PubMed]
  • Maliga P, Sz-Breznovits A, Márton L. Streptomycin-resistant plants from callus culture of haploid tobacco. Nat New Biol. 1973 Jul 4;244(131):29–30. [PubMed]
  • Miesenböck G, Rothman JE. The capacity to retrieve escaped ER proteins extends to the trans-most cisterna of the Golgi stack. J Cell Biol. 1995 Apr;129(2):309–319. [PMC free article] [PubMed]
  • Munro S, Pelham HR. A C-terminal signal prevents secretion of luminal ER proteins. Cell. 1987 Mar 13;48(5):899–907. [PubMed]
  • Navazio L, Baldan B, Mariani P, Gerwig GJ, Vliegenthart JF. Primary structure of the N-linked carbohydrate chains of Calreticulin from spinach leaves. Glycoconj J. 1996 Dec;13(6):977–983. [PubMed]
  • Nishimura N, Balch WE. A di-acidic signal required for selective export from the endoplasmic reticulum. Science. 1997 Jul 25;277(5325):556–558. [PubMed]
  • Pahl HL, Baeuerle PA. Endoplasmicreticulum-induced signal transduction and gene expression. Trends Cell Biol. 1997 Feb;7(2):50–55. [PubMed]
  • Pedrazzini E, Giovinazzo G, Bielli A, de Virgilio M, Frigerio L, Pesca M, Faoro F, Bollini R, Ceriotti A, Vitale A. Protein quality control along the route to the plant vacuole. Plant Cell. 1997 Oct;9(10):1869–1880. [PMC free article] [PubMed]
  • Pelham HR. Evidence that luminal ER proteins are sorted from secreted proteins in a post-ER compartment. EMBO J. 1988 Apr;7(4):913–918. [PMC free article] [PubMed]
  • Pelham HR. Sorting and retrieval between the endoplasmic reticulum and Golgi apparatus. Curr Opin Cell Biol. 1995 Aug;7(4):530–535. [PubMed]
  • Pueyo JJ, Chrispeels MJ, Herman EM. Degradation of transport-competent destabilized phaseolin with a signal for retention in the endoplasmic reticulum occurs in the vacuole. Planta. 1995;196(3):586–596. [PubMed]
  • Rabouille C, Hui N, Hunte F, Kieckbusch R, Berger EG, Warren G, Nilsson T. Mapping the distribution of Golgi enzymes involved in the construction of complex oligosaccharides. J Cell Sci. 1995 Apr;108(Pt 4):1617–1627. [PubMed]
  • Scheel J, Pepperkok R, Lowe M, Griffiths G, Kreis TE. Dissociation of coatomer from membranes is required for brefeldin A-induced transfer of Golgi enzymes to the endoplasmic reticulum. J Cell Biol. 1997 Apr 21;137(2):319–333. [PMC free article] [PubMed]
  • Spang A, Schekman R. Reconstitution of retrograde transport from the Golgi to the ER in vitro. J Cell Biol. 1998 Nov 2;143(3):589–599. [PMC free article] [PubMed]
  • Tatu U, Helenius A. Interactions between newly synthesized glycoproteins, calnexin and a network of resident chaperones in the endoplasmic reticulum. J Cell Biol. 1997 Feb 10;136(3):555–565. [PMC free article] [PubMed]
  • Townsley FM, Wilson DW, Pelham HR. Mutational analysis of the human KDEL receptor: distinct structural requirements for Golgi retention, ligand binding and retrograde transport. EMBO J. 1993 Jul;12(7):2821–2829. [PMC free article] [PubMed]
  • Vitale A, Denecke J. The endoplasmic reticulum-gateway of the secretory pathway . Plant Cell. 1999 Apr;11(4):615–628. [PMC free article] [PubMed]
  • Vollenweider F, Kappeler F, Itin C, Hauri HP. Mistargeting of the lectin ERGIC-53 to the endoplasmic reticulum of HeLa cells impairs the secretion of a lysosomal enzyme. J Cell Biol. 1998 Jul 27;142(2):377–389. [PMC free article] [PubMed]
  • Wandelt CI, Khan MR, Craig S, Schroeder HE, Spencer D, Higgins TJ. Vicilin with carboxy-terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants. Plant J. 1992 Mar;2(2):181–192. [PubMed]

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