Abstract
Using in vivo pulse labeling, changes in the pattern of protein synthesis were detected in isolated barley aleurone layers treated with fibberellic acid (GA3). GA3 greatly altered the relative rates of synthesis of many polypeptides, increasing some, notably α-amylase, and decreasing others. α-Amylase synthesis increased until it was the major product (over 60%) of protein synthesis after 24h. The pulse-labeled pattern of secreted polypeptides was also changed by GA3. There was the expected increase in α-amylase together with a number of other polypeptides but there was reduced secretion of several polypeptides also.
Cell-free translation of RNA isolated from control and hormone-treated tissues was used to measure changes in mRNA levels. GA3 caused many changes, particularly in the level of mRNA for α-amylase. In vitro synthesized α-amylase, identified by immunoaffinity chromatography, had an Mr of 46 000. This polypeptide was partially processed to a polypeptide with Mr 44 000 by the addition of dog pancreas membranes to the in vivo translation mixture. The level of mRNA for α-amylase began to increase 2–4 h after GA3 was added and reached a maximum level of about 20% of total mRNA after 16 h. Thus after 16 h, the synthesis of α-amylase as a proportion of total protein synthesis, continued to increase while the level of its mRNA as a proportion of total mRNA remained constant. These results indicate that protein synthesis was modified more extensively than we can account for by changes in mRNA.
Abscisic acid (ABA) reversed all of the effects of GA3 on protein synthesis and mRNA levels. It also promoted synthesis of a small number of new polypeptides and increased the level of some mRNAs. GA3 reversed the accumulation of ABA-promoted mRNAs. Although, ABA strongly suppressed the increase in the level of translatable mRNA for α-amylase, there was an even stronger inhibition of enzyme synthesis and accumulation.
We conclude that both GA3 and ABA regulate protein synthesis both positively and negatively in aleurone cells largely by regulating levels of mRNA and in the case of α-amylase, possibly also by changing the efficiency of translation of its mRNA.
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Andersson K, Benyamin Y, Douzou P and Balny C (1978) Organic solvents and temperature effects on desorption from immunoadsorbents. DNP-BSA anti-DNP as a model. J Immunol Meth 23, 17–21
Bennett PA and Chrispeels MJ (1972) De novo synthesis of ribonuclease and β-1, 3-glucanase by aleurone cells of barley. Plant Physiol 49, 445–447
Bernal-Lugo I, Beachy RN and Varner JE (1981) The response of barley aleurone layers to gibberellic acid includes the transcription of new sequences. Biochem Biophys Res Commun 102, 617–623
Blobel G and Dobberstein B (1975) Transfer of proteins across membranes I Presence of proteolytically processed and unprocessed nascent immunoglobin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol 67, 835–851
Chrispeels MJ and Varner JE (1966) Inhibition of gibberellic acid induced formation of α-amylase by abscisin II. Nature 212, 1066–1067
Chrispeels MJ and Varner JE (1967) Gibberellic acid-enhanced synthesis and release of α-amylase and ribonuclease by isolated barley aleurone layers. Plant Physiol 42, 398–406
Chrispeels MJ and Varner JE (1967) Hormonal control of enzyme synthesis: On the mode of action of gibberellic acid and abscisin in aleurone layers of barley. Plant Physiol 42, 1008–1016
Ferrari TE and Varner JE (1969) Substrate induction of nitrate reductase in barley aleurone layers. Plant Physiol 44, 85–88
Filner P and Varner JE (1967) A test for de novo synthesis of enzymes: density labelling with H2 18O of barley α-amylase induced by gibberellic acid. Proc Nat Acad Sci (Wash) 58, 1520–1526
Gerace L and Blobel G (1980) The nuclear envelope lamina is reversibly depolymerized during mitosis. Cell 19, 277–287
Gill DS, Kumarasamy R and Symons RH (1981) Cucumber mosaic virus-induced RNA replicase: Solubilization and partial purification of the particulate enzyme. Virol 113, 1–8
Hardie DG (1975) Control of carbohydrate formation by gibberellic acid in barley endosperm. Phytochem 14, 1719–1722
Higgins TJV and Jacobsen JV (1978) Phytohormones and subcellular structural modification. In Letham DS, Goodwin PB and Higgins TJV eds. Phytohormones and related compounds: A comprehensive treatise Vol. 1. The Biochemistry of of phytohormones and related compounds, pp 419–465. Elsevier/North Holland, Amsterdam, Oxford, New York
Higgins TJV and Jacobsen JV (1978) The influence of plant hormones on selected aspects of cellular metabolism. In Letham DS, Goodwin PB and Higgins TJV eds. Phytohormones and related compounds, pp 467–514. Elsevier/North Holland, Amsterdam, Oxford, New York
Higgins TJV and Spencer D (1977) Cell-free synthesis of pea seed proteins. Plant Physiol 60, 655–661
Higgins TJV and Spencer D (1981) Precursor forms of pea vicilin subunits: modification by microsomal membranes during cell-free translation. Plant Physiol 67, 205–211
Higgins TJV, Zwar JA and Jacobsen JV (1976) Gibberellic acid enhances the level of translatable mRNA for α-amylase in barley aleurone layers. Nature 260, 166–169
Higgins TJV, Zwar JA and Jacobsen JV (1977) Hormonal control of the level of translatable mRNA for α-amylase in barley aleurone layers. In Acides nucleiques et synthese de proteines chez les vegetaux, pp 481–486. Coll. Internat. No 261, Paris, Centre National Recherche Sci
Ho DT-H and Varner JE (1974) Hormonal control of messenger ribonucleic acid metabolism in barley aleurone layers. Proc Natl Acad Sci USA 71, 4783–4786
Ho DT-H and Varner JE (1976) Response of barley aleurone layers to abscisic acid. Plant Physiol 57, 175–178
Ho DT-H (1979) Hormonal control of enzyme formation in barley aleurone layers. In Rubinstein I, Phillips RL, Green CE and Gegenback BC, eds. Plant molecular biology, pp 217–240. New York: Academic Press
Ho DT-H (1980) Hormonal and genetic regulation of α-amylase synthesis in barley aleurone cells. In Leaver CJ. ed. Genome organization and expression in plants, pp 147–157. New York and London, Plenum Press
Jacobsen JV (1977) Regulation of ribonucleic acid metabolism by plant hormones. Annu Rev Plant Physiol 28, 537–564
Jacobsen JV and Higgins TJV (1978) The influence of phytohormones on replication and transcription. In Letham DS, Goodwin PB and Higgins TJV eds. Phytohormones and related compounds: A comprehensive treatise. Vol. 1. The biochemistry of phytohormones and related compounds, pp 515–582. Elsevier/North Holland, Amsterdam, Oxford, New York
Jacobsen JV and Higgins TJV (1978) Posttranscriptional, translational and posttranslational effects of plant hormones. In Letham DS, Goodwin PB and Higgins TJV eds. Phytohormones and related compounds: A comprehensive treatise. Vol. 1. The biochemistry of phytohormones and related compounds, pp 583–621. Elsevier/North Holland, Amsterdam, Oxford, New York
Jacobsen JV and Varner JE (1967) Gibberellic acid induced synthesis of protease by isolated aleurone layers of barley. Plant Physiol 42, 1595–1600
Jacobsen JV, Higgins TJV and Zwar JA (1979) Hormonal control of endosperm function during germination. In Rubenstein I, Phillips R, Green C, and Gegenbach B, eds. The plant seed-development, preservation and germination, pp 241–262. New York, Academic Press
Johnson KD and Kende H (1971) Hormonal control of lecithin synthesis in barley aleurone cells: regulation of CDP-choline pathway by gibberellin. Proc Natl Acad Sci USA 68, 2674–2677
Johnson KD and Chrispeels MJ (1973) Regulation of pentosan biosynthesis in barley aleurone tissue by gibberellic acid. Planta 111, 353–364
Jones RL and Jacobsen JV (1978) Membrane and RNA metabolism in the response of aleurone cells to GA. Bot Mag Tokyo Special Issue 1, 83–99
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277, 680–685
Lowry OH, Rosebrough NJ, Farr AL and Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193, 265–275
Mans RJ and Novelli GD (1961) Measurement of the incorporation of radioactive amino acids into protein by a filter-paper disc method. Arch Biochem Biophys 94, 48–53
Mozer TJ (1980) Control of protein synthesis in barley aleurone layers by the plant hormones gibberellic acid and abscisic acid. Cell 20, 479–485
Mutukrishnan S, Chandra GR and Maxwell ES (1979) Hormone-induced increase in levels of functional mRNA and α-amylase mRNA in barley aleurones. Proc Natl Acad Sci (USA) 76, 6181–6185
Okita TW, deCaleys R and Rappaport L (1979) Synthesis of a possible precursor of α-amylase in wheat aleurone cells. Plant Physiol 63, 195–200
Shields D and Blobel G (1978) Efficient cleavage and segregation of nascent presecretory proteins in a reticulocyte lysate supplemented with microsomal membranes. J Biol Chem 253, 3753–3756
Silvanovich MP and Hill RD (1976) Affinity chromatography of cereal α-amylase. Anal Biochem 73, 430–433
Spencer D, Higgins TJV, Button SC and Davey RA (1980) Pulse-labeling studies on protein synthesis in developing pea seeds and evidence of a precursor form of legumin small subunit. Plant Physiol 66, 510–515
Ternynck T and Avrameus S (1976) Polymerization and immobilization of proteins using ethylchloroformate and glutaraldehyde. Scand J Immunol Suppl 3, 29–35
Varner JE and Chandra GR (1964) Hormonal control of enzyme synthesis in barley endosperm. Proc Natl Acad Sci USA 52, 100–106
Varner JE and Ho DT (1976) The role of hormones in the integration of seedling growth. In Papaconstantinou J, ed. The molecular biology of hormone action, pp 173–194. New York, San Francisco and London: Academic Press
Walton DC (1980) Biochemistry and physiology of abscisic acid. Annu Rev Plant Physiol 31, 453–489
Zwar JA and Jacobsen JV (1972) A correlation between a ribonucleic acid fraction selectively labeled in the presence of gibberellic acid and amylase synthesis in barley aleurone layers. Plant Physiol 49, 1000–1006
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Higgins, T.J.V., Jacobsen, J.V. & Zwar, J.A. Gibberellic acid and abscisic acid modulate protein synthesis and mRNA levels in barley aleurone layers. Plant Mol Biol 1, 191–215 (1982). https://doi.org/10.1007/BF00021032
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DOI: https://doi.org/10.1007/BF00021032