Abstract
The growth hormone gene of the rainbow trout (tGH-II) was expressed in leaves of transgenic tobacco plants and seeds of transgenicArabidopsis plants using tissue-specific promoters. Although in the leaves and the seeds comparble amounts of tGH-II mRNA could be detected, the protein could only be identified in the tobacco leaves. Passage of the hormone into the secretory pathway, mediated by the signal sequence of the extracellular tobacco PRI-b (pathogenesis-related) protein, resulted in correct disulphide bridge formation and (partial) glycosylation of the hormone. In contrast, cytoplasmic expression resulted in misfolding and partial breakdown of the protein. The data demonstrate that synthesis, folding and glycosylation of heterologous proteins in plants is dependent both on subcellular location as well as on the tissue or cell type in which the protein is expressed.
Similar content being viewed by others
References
Bednarek, S.Y. and Raikhel, N.V. (1991) The barley lectin carboxyl terminal propeptide is a vacuolar protein sorting determinant in plants.Pl. Cell 3, 1195–206.
Cornelissen, M. and Vandewiele, M. (1989) Nuclear transcriptional activity of the tobacco plastidpbsA promoter.Nucl. Acids Res. 17, 19–29.
De Almeida, E.R.P., Gossele, V., Muller, C.G., Dockx, J., Reynaerts, A., Botterman, J., Krebbers, E. and Timko, M.P. (1989) Transgenic expression of two marker genes under the control of anArabidopsis rbcS promoter: sequences encoding the Rubisco transit peptide increase expression levels.Mol. Gen. Genet. 218, 78–86.
Denecke, J., Gossele, V., Botterman, J. and Cornelissen, M. (1989) Quantitative analysis of transiently expressed genes in plant cells.Meth. Mol. Cell. Biol. 1, 19–27.
Denecke, J., Botterman, J. and Deblaere, R. (1990) Protein secretion in plant cells can occur via a default pathway.Pl. Cell 2, 51–9.
De Neve, M., De Loose, M., Jacobs, A., Van Houdt, H., Kaluza, B., Weidle, U., Van Montagu, M. and Depicker, A. (1993) Assembly of an antibody and its derived antibody fragment inNicotiana andArabidopsis.Transgenic Res. 2, 227–37.
De Zoeten, G.A., Penswick, J.R., Horisberger, M.A. Ahl, P., Schultze, M. and Hohn, T. (1989) The expression, localization and effect of a human interferon in plants.Virology 172, 213–22.
Düring, K., Hippe, S., Kreuzaler, F. and Schell, J. (1990) Synthesis and self-assembly of a functional monoclonal antibody in transgenicNicotiana tabacum.Pl. Mol. Biol. 15, 281–93.
Faye, L., Johnson, K.D., Sturm, A. and Chrispeels, M.J. (1989) Structure, biosynthesis, and function of asparagine-linked glycans on plant glycoproteins.Physiologia Plantarum 75, 309–14.
Gething, M.J. and Sambrook, J. (1992) Protein folding in the cell.Nature 355, 33–45.
Gielen, J., De Beuckeleer, M., Seurinck, J., Deboeck, F., De Greve, H., Lemmers, M., Van Montagu, M. and Schell, J. (1984) The complete nucleotide sequence of the T l -DNA of theAgrobacterium tumefaciens plasmid pTiAch5.EMBO J. 3, 835–46.
Grill, L., Erwin, R.L., Berliner, D.L. and Hubbard, E.T. (1989) PCT patent application WO 8908145.
Guerche, P., Tire, C., Grossi De Sa, F., De Clercq, A., Van Montagu, M. and Krebbers, E. (1990) Differential expression of theArabidopsis 2S albumin genes and the effect of increasing gene family size.Pl. Cell 2, 469–78.
Hiatt, A., Cafferkey, R. and Bowdish, K. (1989) Production of antibodies in transgenic plants,Nature 342, 76–78.
Horsch, R.B., Fry, J.E., Hoffman, N.L., Eichholtz, D., Rogers, S.G. and Fraley, R.T. (1985) A simple and general method for transferring genes into plants.Science 227, 1229–31.
Hunt, D.C. and Chrispeels, M.J. (1991) The signal peptide for vacuolar protein is necessary and sufficient for the efficient secretion of cytosolic protein.Pl. Physiol. 96, 18–25.
Josephson, S. and Bishop, R. (1988) Secretion of peptides fromE. Coli: a production system for the pharmaceutical industry. Trends Biotech.6, 218–24.
Krebbers, E., Bosch, D. and Vandekerckhove, J. (1992). Prospects and progress in the production of foreign proteins and peptides in plants. In: Shewry, P. and Gutteridge, S. edsPlant Protein Engineering, pp. 315–325. Cambridge, UK: Cambridge University Press.
Le Bail, P.Y., Sumpter, J.P., Carragher, J., Mourot, B., Nur, P.D. and Weil, C. (1991) Development and validation of a highly sensitive radioimmunoassay to chinook salmon (Onchorynchus tshawytscha) growth hormone.Gen. Comp. Endocrinol. 83, 75–85.
Matsuoka, K. and Nakamura, K. (1991) Propeptide of a precursor to a plant vacuolar protein required for vacuolar targeting.Proc. Natl Acad. Sci. USA 88, 834–8.
Rentier-Debrue, F., Swennen, D., Mercier, L., Lion, M., Benrubi, O. and Martial, J. (1989) Molecular cloning and characterization of two forms of trout growth hormone cDNA: expression and secretion of tGH-II byEscherichia coli.DNA 8, 109–17.
Sijmons, P.C., Dekker, B.M.M., Schrammeijer, B., Verwoerd, T.C., Van den Elzen, P.J.M. and Hoekema, A. (1990) Production of correctly processed human serum albumin in transgenic plants.Bio/Technology 8, 217–21.
Tarentino, A.L., Gomez, C.M. and Plummer, T.H., Jr. (1985) Deglycosylation of asparagine-linked glycans by peptide: N-glycosidase F.Biochemistry 24, 4665–71.
Valvekens, D., Van Montagu, M. and Van Lijsebettens, M. (1988)Agrobaterium tumefaciens mediated transformation ofArabidopsis root explants using kanamycin selection.Proc. Natl Acad. Sci. USA 85, 5536–40.
Van Brunt, J. (1988) Molecular farming: transgenic animals as bioreacctors.Bio/Technology 6, 1149–54.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bosch, D., Smal, J. & Krebbers, E. A trout growth hormone is expressed, correctly folded and partially glycosylated in the leaves but not the seeds of transgenic plants. Transgenic Research 3, 304–310 (1994). https://doi.org/10.1007/BF01973590
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01973590