Abstract
To demonstrate the relationship between protein-bodies in seeds and vacuoles in other tissues, we expressed the coding sequences of two bean (Phaseolus vulgaris L.) protein-body proteins in tobacco (Nicotiana tabacum L.). We chose phytohemagglutinin-L (PHA-L) and tonoplast intrinsic protein (TIP) as representatives of the protein-body contents and protein-body membrane, respectively. The location of the two proteins in the leaves of transgenic tobacco was examined by immunocytochemistry and in preparations of isolated vacuoles. Tonoplast intrinsic protein accumulates primarily in tonoplasts in tobacco leaves, whereas PHA is found exclusively in the vacuolar sap, showing that the signals that target proteins to protein-bodies and their limiting membranes in seeds are correctly recognized in leaves. This observation provides further evidence that proteinbodies of dicotyledonous seeds should be considered as protein-storage vacuoles.
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Abbreviations
- TIP:
-
tonoplast intrinsic protein
- PHA:
-
phytohemagglutinin
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
References
Barker, R.F., Idler, K.B., Thompson, D.V., Kemp, J.D. (1983) Nucleotide sequence of the T-DNA region of Agrobacterium tumefaciens octopine Ti plasmid pTi15955. Plant Mol. Biol. 2, 335–350
Boller, T., Kende, H. (1979) Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 63, 1123–1132
Craig, S., Goodchild, D.J., Miller, C. (1980) Structural aspects of protein accumulation in developing pea cotyledons. II. Threedimensional reconstructions of vacuoles and protein bodies from serial sections. Aust. J Plant Physiol. 7, 329–337
Damm, B., Willmitzer, L. (1988) Regeneration of fertile plants from protoplasts of different Arabidopsis thaliana genotypes. Mol. Gen. Genet. 213, 15–20
Denecke, J., Botterman, J., Deblaere, R. (1990) Protein secretion in plant cells can occur via a default pathway. Plant Cell 2, 51–59
Greenwood, J.S., Chrispeels, M.J. (1985) Correct targeting of the bean storage protein phaseolin in the seeds of transformed tobacco. Plant Physiol. 79, 65–71
Guerrero, F.D., Jones, J.T., Mullet, J.E. (1990) Turgor-responsive gene transcription and RNA levels increase rapidly when pea shoots are wilted. Sequence and expression of three inducible genes. Plant Mol. Biol. 15, 11–26
Herman, E.M., Baumgartner B., Chrispeels, M.J. (1981) Uptake and apparent digestion of cytoplasmic organelles by protein bodies (protein storage vacuoles) in mung bean cotyledons. Eur. J. Cell Biol. 24, 226–235
Herman, E.M., Hankin, C.N., Shannon, L.M. (1988) Bark and leaf lectins of Sophora japonica are sequestered in protein-storage vacuoles. Plant Physiol. 86, 1027–1031
Higgins, T.J.V., Newbigin, E.J., Spencer, D., Llewellyn, D.J., Craig, S. (1989) The sequence of a pea vicilin gene and its expression in transgenic tobacco plants. Plant Mol. Biol. 11, 683–695
Höfgen, R., Willmitzer, L. (1988) Storage of competent cells for Agrobacterium transformation. Nucl. Acids Res. 16, 9877
Johnson, K.D., Herman, E.M., Chrispeels, M.J. (1989) An abundant, highly conserved tonoplast protein in seeds. Plant Physiol. 91, 1006–1013
Johnson, K.D., Höfte, H., Chrispeels, M.J. (1990) An intrinsic tonoplast protein of protein storage vacuoles in seeds is structurally related to a bacterial solute transporter (Glpf). Plant Cell 2, 525–532
Lowry, O.H., Rosebrough, N.J., Farr A.L., Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275
Matile, P. (1975) The lytic compartment of plant cells. Springer-Verlag, New York
Mettler, I.J., Leonard, R.T. (1979) Isolation and partial characterization of vacuoles from tobacco protoplasts. Plant Physiol. 64, 1114–1120
Öpik, H. (1968) Development of cotyledon cell structure in ripening Phaseolus vulgaris seeds. J. Exp. Bot. 19, 64–76
Pietrzak, M., Shillito, R.D., Hohn, T., Potrykus, I. (1986) Expression in plants of two bacterial antibiotic resistance genes after protoplast transformation with a new plant expression vector. Nucl. Acids Res. 14, 5857–5868
Sonnewald, U., Sturm, A., Chrispeels, M.J., Willmitzer, L. (1989) Targeting and glycosylation of patatin, the major potato tuber protein in leaves of transgenic tobacco. Planta 179, 171–180
Staswick, P.E. (1990) Novel regulation of vegetative storage protein genes. Plant Cell 2, 1–6
Sturm, A., Voelker, T.A., Herman, E.M., Chrispeels, M.J. (1988) Correct glycosylation, Golgi-processing and targeting to protein bodies of the vacuolar protein phytohemagglutinin in transgenic tobacco. Planta 175, 170–183
Tague, B.W., Chrispeels, M.J. (1987) The plant vacuolar protein, phytohemagglutinin, is transported to the vacuoles of transgenic yeast. J. Cell Biol. 105, 1971–1979
Van der Wilden, W., Herman, E.M., Chrispeels, M.J. (1980) Protein bodies of mung bean cotyledons as autophagic organelles. Proc. Natl. Acad. Sci. USA 77, 428–432
Voelker, T.A., Florkiewicz, R.Z., Chrispeels, M.J. (1986) Secretion of phytohemagglutinin by monkey COS cells. Eur. J. Cell Biol. 42, 218–223
Voelker, T., Sturm, A., Chrispeels, M.J. (1987) Differences in expression between two seed lectin alleles obtained from normal and lectin-deficient beans are maintained in transgenic tobacco. EMBO J. 6, 3571–3577
Voelker, T.A., Herman, E.M., Chrispeels, M.J. (1989) In vitro mutated phytohemagglutinin genes expressed in tobacco seeds: role of glycans in protein targeting and stability. Plant Cell 1, 95–104
Wetzel, S., Demmers, C., Greenwood, J.S. (1989) Seasonally fluctuating bark proteins are a potential form of nitrogen storage in three temperate hardwoods. Planta 178, 275–281
Wilkins, T.A., Bednarek, S.Y., Raikhel, N.V. (1990) Role of propeptide glycan in post-translational processing and transport of barley lectin to vacuoles in transgenic tobacco. Plant Cell 2, 301–313
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This work has been supported by a grant from the U.S. Department of Agriculture Competitive Research Grants Office to Maarten J. Chrispeels. Herman Höfte is a recipient of a Postdoctoral Fellow-ship from the European Molecular Biology Organization. Craig Dickinson is a recipient of a National Institutes of Health Postdoctoral Fellowship. Loîc Faye was on leave from the Centre National de la Recherche Scientifique, UA203, Université de Rouen, Mont Saint Aignan France and supported by a cooperative CNRS — National S cience Foundation grant. The mention of vendor or product in this paper does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over vendors of similar products not mentioned. We thank Jürgen Denecke for providing plasmid pDE1001, Antje von Schaewen for the plant expression cassette and Marie-Theres Hauser for the modified vacuole preparation protocol.
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Höfte, H., Faye, L., Dickinson, C. et al. The protein-body proteins phytohemagglutinin and tonoplast intrinsic protein are targeted to vacuoles in leaves of transgenic tobacco. Planta 184, 431–437 (1991). https://doi.org/10.1007/BF00197889
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DOI: https://doi.org/10.1007/BF00197889