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Transcription and translation of phloem protein (PP2) during phloem differentiation in Cucurbita maxima

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Abstract

The synthesis of a major phloem protein, PP2, was investigated by measurement of the mRNA at various stages of phloem development in Cucurbita. Quantitative assays with immuno-electrophoresis showed that the amounts of PP2 in hypocotyls of Cucurbita seedlings increased with the age of seedlings. An increase in mRNA for PP2 during the early stages of seedling growth was also observed by immunoprecipitation of the invitro translation products of hypocotyl polyadenylated RNA. There was close timing in the variations of PP2 synthesised in vivo and in the changes in amounts of translatable PP2-mRNA during the course of seedling growth. A complementary-DNA (cDNA) library to polyadenylated RNA from hypocotyls of 3-d-old Cucurbita seedlings has been constructed. Two cDNA clones, A and B, have been identified by hybrid-release translation to be complementary to the mRNA coding for PP2. The levels of total mRNA for PP2 measured with clone A were found to increase in the first 4 d of seedling growth but decreased to lower levels in older seedlings. Regulatory controls on both transcription and modification of transcripts appeared to occur during the synthesis of PP2.

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Abbreviations

cDNA:

complementary DNA

(GlcNAc)4 :

tetra-(2-acetamido-2-deoxy-β-d-glucopyranoside)

Mr :

relative molecular mass

PP1:

phloem protein 1

PP2:

phloem protein 2

poly(A)+RNA:

polyadenylated RNA

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

References

  • Allen, A.K. (1979) A lectin from the exudate of the fruit of the vegetable marrow (Cucurbita pepo) that has a specificity for β-1,4-linked N-acetylglucosamine oligosaccharides. Biochem. J. 183, 133–137

    Google Scholar 

  • Alosi, M.C., Park, R.B. (1983) Fractionation and polypeptide analysis of phloem tissue of Pinus sabiniana Dougl. Planta 157, 298–306

    Google Scholar 

  • Beyenbach, J., Weber, C., Kleinig, H. (1974) Sieve-tube proteins from Cucurbita maxima. Planta 119, 113–124

    Google Scholar 

  • Carrington, D.M., Auffret, A., Hanke, D.E. (1985) Polypeptide ligation occurs during post-translational modification of concanavalin A. Nature 313, 64–67

    Google Scholar 

  • Cronshaw, J. (1975) P-proteins. In: Phloem transport, pp. 79–147, Aronoff, S., Dainty, J., Gorham, P.R., Srivastave, L.M., Swanson, C.A., eds. Plenum Press, New York

    Google Scholar 

  • Esau, K., Cronshaw, J. (1967) Tubular components in cells of healthy and tobacco mosaic virus-infected Nicotiana. Virology 33, 26–35

    Google Scholar 

  • Gietl, C., Ziegler, H. (1979) mRNA in phloem exudates. Naturwissenschaften 66, 523–524

    Google Scholar 

  • Gietl, C., Ziegler, H. (1980) Affinity chromatoraphy of carbohydrate binding proteins in the phloem exudate from several tree species. Biochem. Physiol. Pflanzen 175, 50–57

    Google Scholar 

  • Grunstein, M., Hogness, D.S. (1975) Colony hybridization: A method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad. Sci. USA 72, 3961–3965

    Google Scholar 

  • Gubler, U., Hoffman, B.J. (1983) A simple and very efficient method for generating cDNA libraries. Gene 25, 263–269

    Google Scholar 

  • Hanahan, D. (1983) Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166, 557–580

    Google Scholar 

  • Johnson, R.P.C. (1978) The microscopy of P-protein filaments in freeze-etched sieve pores. Brownian motion limits resolution of their positions. Planta 143, 191–205

    Google Scholar 

  • Kleinig, M., Thomes, J., Dorr, I., Kollmann, R. (1975) Filament formation in vitro of a sieve tube protein from Cucurbita maxima and Cucurbita pepo. Planta 127, 163–170

    Google Scholar 

  • Kollmann, R., Dorr, I., Kleinig, H. (1970) Protein filaments-structural components of the phloem exudate. I. Observations with Cucurbita and Nicotiana. Planta 95, 86–94

    Google Scholar 

  • Laurell, C.-B. (1966) Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal. Biochem. 15, 45–52

    Google Scholar 

  • Maniatis, T., Fritsch, E.F., Sambrook, J. (1982) Molecular cloning — a laboratory manual. pp. 86–94, Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Martin, C., Northcote, D.H. (1981) Qualitative and quantitative changes in mRNA of castor beans during the initial stages of germination. Planta 151, 189–197

    Google Scholar 

  • Martin, C., Northcote, D.H. (1982) The action of exogenous gibberellic acid on isocitrate lyase-mRNA in germinating castor bean seeds. Planta 154, 174–183

    Google Scholar 

  • McEuen, A.R., Hill, H.A.O. (1982) Superoxide, hydrogen peroxide, and the gelling of phloem sap from Cucurbita pepo. Planta 154, 295–297

    Google Scholar 

  • Nuske, J., Eschrich, W. (1976) Synthesis of P-protein in mature phloem of Cucurbita maxima. Planta 132, 109–118

    Google Scholar 

  • Palevitz, B.A., Newcomb, E.H. (1971) The ultrastructure and development of tubular and crystalline P-protein in the sieve elements of certain papilionaceous legumes. Protoplasma 72, 399–426

    Google Scholar 

  • Parnes, J.R., Velan, B., Felsenfeld, A., Ramanathan, L., Ferrini, U., Appella, E., Seidman, J.G. (1981) Mouse β2-microglobulin cDNA clones: A screening procedure for cDNA clones corresponding to rare mRNAs. Proc. Natl. Acad. Sci. USA 78, 2253–2257

    Google Scholar 

  • Pelham, H.R.B., Jackson, R.J. (1976) An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. J. Biochem. 67, 247–256

    Google Scholar 

  • Read, S.M., Northcote, D.H. (1981) Minimization of variation in the response to different proteins of the Coomassie Blue G dye-binding assay for protein. Anal. Biochem. 116, 53–64

    Google Scholar 

  • Read, S.M., Northcote, D.H. (1983a) Subunit structure and interactions of the phloem proteins of Cucurbita maxima (pumpkin). Eur. J. Biochem. 134, 561–569

    Google Scholar 

  • Read, S.M., Northcote, D.H. (1983b) Chemical and immunological similarities between the phloem proteins of three genera of the Cucurbitaceae. Planta 158, 119–127

    Google Scholar 

  • Rosbash, M., Ford, P.J. (1974) Polyadenylic acid-containing RNA in Xenopus laevis oocytes. J. Mol. Biol. 85, 87–101

    Google Scholar 

  • Sabnis, D.D., Hart, J.W. (1976) A comparative analysis of phloem exudate proteins from Cucumis melo, Cucumis sativus and Cucurbita maxima by polyacrylamide gel electrophoresis and isoelectric focussing. Planta 130, 211–218

    Google Scholar 

  • Sabnis, D.D., Hart, J.W. (1979) Heterogeneity in phloem protein complements from different species. Consequences to hypotheses concerned with P-protein function. Planta 145, 459–466

    Google Scholar 

  • Sloan, R.T., Sabnis, D.D., Hart, J.W. (1976) The heterogeneity of phloem exudate proteins from different plants: A comparative survey of ten plants using polyacrylamide gel electrophoresis. Planta 132, 97–102

    Google Scholar 

  • Smith, L.M., Sabnis, D.D. (1968) Immunocytochemistry of phloem lectins. In: Phloem transport (Plant Biology Series, vol. I) pp. 157–159, Cronshaw, J., Lucas, W.J., Giaquinta, R.T., eds. Alan R. Liss, New York

    Google Scholar 

  • Thomas, P.S. (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. USA 77, 5201–5205

    Google Scholar 

  • Walker, T.S. (1972) The purification and some properties of a protein causing gelling in phloem sieve tube exudate from Cucurbita pepo. Biochim. Biophys. Acta 257, 433–444

    Google Scholar 

  • Walker, T.S., Thaine, R. (1971) Proteins and fine structural components in exudate from sieve tubes in Cucurbita pepo stems. Ann. Bot. 35, 773–790

    Google Scholar 

  • Weber, C., Franke, W.W., Kartenbeck, J. (1974) Structure and biochemistry of phloem-proteins isolated from Cucurbita maxima. Exp. Cell Res. 87, 79–106

    Google Scholar 

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Sham, MH., Northcote, D.H. Transcription and translation of phloem protein (PP2) during phloem differentiation in Cucurbita maxima . Planta 170, 392–399 (1987). https://doi.org/10.1007/BF00395032

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  • DOI: https://doi.org/10.1007/BF00395032

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