Abstract
The increase in extractable phenylalanine ammonia-lyase (PAL;EC 4.3.1.5.) activity induced in French bean cell suspension cultures in response to treatment with autoclaved ribonuclease A was inhibited by addition of the phenylpropanoid pathway intermediates cinnamic acid, 4-coumaric acid or ferulic acid. The effectiveness of inhibition was in the order cinnamic acid>4-coumaric acid>ferulic acid. Cinnamic acid also inhibited the PAL activity increase induced by dilution of the suspensions into an excess of fresh culture medium. Addition of low concentrations (<10-5M) of the pathway intermediates to cultures at the time of application of ribonuclease gave variable responses ranging from inhibition to 30–40% stimulation of the PAL activity measured at 8 h. Following addition of pathway intermediates to cultures 4–5 h after ribonuclease treatment, rapid increases followed by equally rapid declines in PAL activity were observed. The cinnamic acid-stimulated increase in enzyme activity was unaffected by treatment with cycloheximide at a concentration which gave complete inhibition of the ribonuclease-induced response. However, cycloheximide completely abolished the subsequent decline in enzyme activity. Treatment of induced cultures with α-aminooxy-β-phenylpropionic acid (AOPPA) resulted in increased but delayed rates of enzyme appearance when compared to controls not treated with the phenylalanine analogue. The results are discussed in relation to current views on the regulation of enzyme levels in higher plants.
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Abbreviations
- AOPPA:
-
α-aminooxy-β-phenylpropionic acid
- PAL:
-
L-phenylalanine ammonia-lyase (EC 4.3.1.5)
- AOA:
-
α-aminooxyacetic acid
References
Amrhein, N. (1979) Novel inhibitors of phenylpropanoid metabolism in higher plants. In: Regulation of secondary product and plant hormone metabolism. FEBS symposium, vol. 55, pp. 173–182, Luckner, M., Schreiber, K. eds. Pergamon, Oxford New York Toronto Sydney Paris Frankfurt
Attridge, T.H., Johnson, C.B., Smith, H. (1974) Density labelling evidence for the phytochrome-mediated activation of phenylalanine ammonia-lyase in mustard cotyledons. Biochim. Biophys. Acta. 343, 446–451
Attridge, T.H., Smith, H. (1973) Evidence for a pool of inactive phenylalanine ammonia-lyase in Cucumis sativus seedlings. Phytochemistry 12, 1569–1574
Betz, B., Schäfer, E., Hahlbrock, K. (1978) Light-induced phenylalanine ammonia-lyase in cell-suspension cultures of Petroselinum hortense. Quantitative comparison of rates of synthesis and degradation. Arch. Biochem. Biophys. 190, 126–135
Dixon, R.A., Bendall, D.S. (1978a) Changes in phenolic compounds associated with phaseollin production in cell suspension cultures of Phaseolus vulgaris. Physiol. Plant Pathol. 13, 283–294
Dixon, R.A., Bendall, D.S. (1978b) Changes in the levels of enzymes of phenylpropanoid and flavonoid synthesis during phaseollin production in cell suspension cultures of Phaseolus vulgaris. Physiol. Plant Pathol. 13, 295–306
Dixon, R.A., Fuller, K.W. (1976) Effects of synthetic auxin levels on phaseollin production and phenylalanine ammonia-lyase (PAL) activity in tissue cultures of Phaseolus vulgaris L. Physiol. Plant Pathol. 9, 299–312
Dixon, R.A., Lamb, C.J. (1979) Stimulation of de novo synthesis of L-phenylalanine ammonia-lyase in relation to phytoalexin accumulation in Colletotrichum lindemuthianum elicitor-treated cell suspension cultures of French bean. Biochim. Biophys. Acta 586, 453–465
Durst, F. (1976) The correlation of phenylalanine ammonia-lyase and cinnamic acid-hydroxylase activity changes in Jerusalem artichoke tuber tissues. Planta 132, 221–227
Engelsma, G. (1968) Photoinduction of phenylalanine deaminase in gherkin seedlings. III. Effects of excision and irradiation on enzyme development in hypocotyl segments. Planta 82, 355–368
Engelsma, G. (1979) Inhibition of phenylalanine ammonia-lyase by cinnamic acid derivatives. In: Regulation of secondary product and plant hormone metabolism. FEBS symposium, vol. 55. pp. 163–172, Luckner, M., Schreiber, K., eds. Pergamon, Oxford New York Toronto Sydney Paris Frankfurt
Engelsma, G., van Bruggen, J.M.H. (1971) Ethylene production and enzyme induction in excised plant tissues. Plant Physiol. 48, 94–96
Hahlbrock, K., Ragg, H. (1975) Light-induced changes of enzyme activities in parsley cell suspension cultures. Effects of inhibitors of RNA and protein synthesis. Arch. Biochem. Biophys. 166, 41–46
Hahlbrock, K., Wellman, E. (1973) The light-independent induction of enzymes related to phenylpropanoid metabolism in cell suspension cultures from parsley. Biochim. Biophys. Acta. 304, 702–706
Havir, E.A., Hanson, K.R. (1968) L-Phenylalanine ammonia-lyase. II. Mechanism and kinetic properties of the enzyme from potato tubers. Biochemistry 7, 1904–1914
Johnson, C., Attridge, T., Smith, H. (1975) Regulation of phenylalanine ammonia-lyase synthesis by cinnamic acid. Its implication for the light-mediated regulation of the enzyme. Biochim. Biophys. Acta 385, 11–19
Lamb, C.J. (1977) Trans-cinnamic acid as a mediator of the light-stimulated increase in hydroxycinnamoyl CoA: quinate hydroxycinnamoyl transferase. FEBS Lett. 75, 37–39
Lamb, C.J. (1979) Regulation of enzyme levels in phenylpropanoid biosynthesis: characterization of the modulation by light and pathway intermediates. Arch. Biochem. Biophys. 192, 311–317
Lamb, C.J., Dixon, R.A. (1978) Stimulation of de novo synthesis of L-phenylalanine ammonia-lyase during induction of phytoalexin biosynthesis in cell suspension cultures of Phaseolus vulgaris. FEBS Lett. 94, 277–280
Lamb, C.J., Merritt, T.K., Butt, V.S. (1979) Synthesis and removal of phenylalanine ammonia-lyase activity in illuminated discs of potato tuber parenchyme. Biochim. Biophys. Acta 582, 196–212
Lamb, C.J., Rubery, P.H. (1976) Phenylalanine ammonia-lyase and cinnamic acid 4-hydroxylase: Product repression of the level of enzyme activity in potato tuber discs. Planta 130, 283–290
Leggett-Bailey, J. (1962) In: Techniques in protein chemistry, Ch. 11. Elsevier, Amsterdam
McMahon, D. (1975) Cycloheximide is not a specific inhibitor of protein synthesis in vivo. Plant Physiol. 55, 815–821
Schopfer, P. (1977) Phytochrome control of enzymes. Annu. Rev. Plant Physiol. 28, 223–252
Stafford, H.A. (1974) The metabolism of aromatic compounds. Annu. Rev. Plant Physiol. 25, 459–486
Tong, W-F., Schopfer, P. (1976) Phytochrome-mediated de novo synthesis of phenylalanine ammonia-lyase: an approach using pre-induced mustard seedlings. Proc. Natl. Acad. Sci. USA 73, 4017–4021
Ulrich, B., Amrhein, N. (1978) Induction by light of hydroxycinnamoyl-CoA: quinate hydroxycinnamoyl transferase in buck-wheat (Fagopyrum esculentum Moench): Absence of feed-forward control by trans-cinnamate. Planta 138, 69–71
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Dixon, R.A., Browne, T. & Ward, M. Modulation of L-phenylalanine ammonia-lyase by pathway intermediates in cell suspension cultures of dwarf French bean (Phaseolus vulgaris L.). Planta 150, 279–285 (1980). https://doi.org/10.1007/BF00384656
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DOI: https://doi.org/10.1007/BF00384656