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Calmodulin stimulation of unidirectional calcium uptake by the endoplasmic reticulum of barley aleurone

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Abstract

The role of calmodulin (CaM) in gibberellic acid (GA3)-stimulated Ca2+ uptake was investigated in endomembranes isolated from aleurone cells of barley (Hordeum vulgare L.). Unidirectional Ca2+ -uptake activity of endoplasmic reticulum (ER) was higher in membranes isolated from aleurone layers treated for 16 h with GA3 and Ca2+ compared with those isolated from layers incubated in Ca2+ alone. However, the level of uptake from Ca2+-treated tissue could be stimulated to that of the GA3-treated cells by applying exogenous CaM which increased the V max of the Ca2+ transporter approximately threefold. Calcium uptake in ER from GA3-treated tissue was inhibited by the CaM antagonist W7 in 50% of experiments, whereas the activity in membranes from non-GA3-treated tissue was unaffected. Treatment with GA3 also led to a twofold increase in CaM levels in aleurone layers within 4–6 h, paralleling the time course of the stimulation of Ca2+ uptake and preceding the stimulation of α-amylase secretion. We propose that the elevation of Ca2+ uptake into the ER induced by GA3 may be coordinated and regulated by elevated levels of membrane-associated CaM and this may regulate Ca2+-dependent α-amylase synthesis in the lumen of the ER.

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Abbreviations

CaM:

calmodulin

CCR:

cytochrome-c reductase

EGTA:

ethylene glycol bis (β-aminoethylether)-N,N,N′N′-tetraacetic acid

GA3 :

gibberellic acid

GApp:

Golgi apparatus

IDPase:

inosine diphosphatase

PM:

plasma membrane

RIA:

radioimmunoassay

SR:

sarcoplasmic reticulum

TP:

tonoplast

W5:

N-(6-aminohexyl)-1-naphthalenesulfonamide

W7:

N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide

References

  • Andreev, I.G., Koren'kov, V., Molotkovsky, Y.G. (1990) Calmodulin stimulation of Ca2 +/nH+ antiport across the vacuolar membrane of sugar beet taproot. J. Plant Physiol. 136, 3–7

    Google Scholar 

  • Bowman, E.J., Siebers, A., Altendorf, K. (1988) Bafilomycins: a class of inhibitors of membrane ATPases from microorganisms, animal cells, and plant cells. Proc. Natl. Acad. Sci. USA 85, 7972–7976

    Google Scholar 

  • Brauer, D., Schubert, C., Tsu, S.-I. (1990) Characterization of a Ca2+-translocating ATPase from corn root microsomes. Physiol. Plant. 78, 335–344

    Google Scholar 

  • Buckhout, T.J. (1984) Characterization of a Ca2+ transporter in purified endoplasmic reticulum membrane vesicles from Lepidium sativum. Plant Physiol. 76, 962–967

    Google Scholar 

  • Bush, D.S., Jones, R.L. (1987) Measurement of cytosolic calcium in aleurone protoplasts using Indo-1 and Fura-2. Cell Calcium 8, 455–472

    Google Scholar 

  • Bush, D.S., Jones, R.L. (1988) Cytoplasmic calcium and α-amylase secretion from barley aleurone protoplasts. Eur. J. Cell Biol. 46, 466–469

    Google Scholar 

  • Bush, D.S., Jones, R.L. (1990) Hormonal regulation of Ca2+ transport in microsomal vesicles isolated from barley aleurone layers. In: Calcium in plant growth and development, pp. 60–66, Leonard, R.T., Hepler, P.K., eds. American Society of Plant Physiologists, Bethesda

    Google Scholar 

  • Bush, D.R., Sze, H. (1986) Ca2+-transport in tonoplast and endoplasmic reticulum vesicles isolated from cultured carrot cells. Plant Physiol. 80, 549–555

    Google Scholar 

  • Bush D.S. Biswas, A.K., Jones, R.L. (1989a) Gibberellic acid-stimulated Ca2+ accumulation in the endoplasmic reticulum of barley aleurone: Ca2+ transport and steady state levels. Planta 178, 411–420

    Google Scholar 

  • Bush, D.S., Sticher, L., Van Huystee, R., Wagner, D., Jones, R.L. (1989b) The calcium requirement for stability and enzymatic activity of two isoforms of barley aleurone α-amylase. J. Biol. Chem. 264, 19392–19398

    Google Scholar 

  • Butcher, R.D., Evans, D.E. (1987a) Calcium transport by pea root membranes. I. Purification of membranes and characteristics of uptake. Planta 172, 265–273

    Google Scholar 

  • Butcher, R.D., Evans, D.E. (1987b) Calcium transport by pea root membranes. II. Effects of calmodulin and inhibitors. Planta 172, 273–279

    Google Scholar 

  • Carafoli, E. (1991) Calcium pump of the plasma membrane. Physiol. Rev. 71, 129–153

    Google Scholar 

  • Cheung, W.Y. (1971) Cyclic 3′,5′-nucleotide phosphodiesterase: evidence for and properties of a protein activator. J. Biol. Chem. 246, 2859–2869

    Google Scholar 

  • Colyer, J., Wang, J.H. (1991) Dependence of cardiac sarcoplasmic reticulum calcium pump activity on the phosphorylation status of phospholamban. J. Biol. Chem. 266, 17486–17493

    Google Scholar 

  • Dieter, P., Marmé, D. (1980) Ca2+-transport in mitochondrial and microsomal fractions from higher plants. Planta 150, 1–8

    Google Scholar 

  • Dieter, P., Marmé, D. (1983) The effect of calmodulin and far-red light on the kinetic properties of the mitochondrial Ca2+ transport system from corn. Planta 159, 277–281

    Google Scholar 

  • DuPont F.M., Tanaka, C.K., Hurkman, W.J. (1988) Separation and immunological characterization of membrane fractions from barley roots. Plant Physiol. 86, 717–724

    Google Scholar 

  • Evans, D.E., Briars, S.A., Williams, L.E. (1991) Active Ca2+-transport by plant membranes. J. Exp. Bot. 42, 285–303

    Google Scholar 

  • Gilroy, S., Jones, R.L. (1992) Gibberellic acid and abscisic acid coordinately regulate cytoplasmic calcium and secretory activity in barley aleurone protoplasts. Proc. Natl. Acad. Sci. USA 89, 3591–3595

    Google Scholar 

  • Graf, P., Weiler, E.R. (1989) ATPase-driven transport in sealed plasma membrane vesicles prepared by aqueous two-phase partitioning from leaves of Commelina communis. Physiol. Plant. 79, 469–478

    Google Scholar 

  • Hepler, P.K., Wayne, R.O. (1985) Calcium and plant development. Annu. Rev. Plant Physiol. 36, 397–439

    Google Scholar 

  • Hepler, P.K., Palevitz, B.A., Lancelle, S.A., McCauley, M.M., Lichtscheidl, I. (1990) Cortical endoplasmic reticulum in plants. J. Cell Sci. 96, 355–373

    Google Scholar 

  • Hidaka, H., Sasaki, Y., Tanaka, T., Endo, T., Ohno, S., Fufii, Y., Nagata, T. (1981) N-(6-Aminohexyl)-5-chloro-1-naphthalene-sulfonamide, a calmodulin antagonist, inhibits cell proliferation. Proc. Natl. Acad. Sci. USA 78, 4354–4357

    Google Scholar 

  • Hsieh, W.L., Pierce, W.S., Sze H. (1991) Calcium-pumping ATPases in vesicles from carrot cells — stimulation by calmodulin or phosphatidylserine, and formation of a 120 kD phosphoenzyme. Plant Physiol. 97, 1535–1544

    Google Scholar 

  • Jena, P.K., Reddy, A.S., Poovaiah B.W. (1989) Molecular cloning and sequencing of a cDNA for plant calmodulin: signal-induced changes in the expression of calmodulin. Proc. Natl. Acad. Sci. USA 86, 3644–36488

    Google Scholar 

  • Jones, R.L. (1980) Quantitative and qualitative changes in the endoplasmic reticulum of barley aleurone layers. Planta 150, 70–81

    Google Scholar 

  • Jones, R.L., Bush, D.S. (1991) Gibberellic acid regulates the level of a BiP cognate in the endoplasmic reticulum of barley aleurone. Plant Physiol. 97, 456–459

    Google Scholar 

  • Jones, R.L., Jacobsen J.V. (1982) The role of the endoplasmic reticulum in the synthesis and transport of α-amylase in barley aleurone layers. Planta 156, 421–432

    Google Scholar 

  • Jones, R.L., Jacobsen, J.V. (1991) Regulation of synthesis and transport of secreted proteins in cereal aleurone. Int. Rev. Cytol. 126, 49–88

    Google Scholar 

  • Jones, R.L., Varner, J.E. (1967) The bioassay of gibberellins. Planta 72, 411–420

    Google Scholar 

  • Lew, R.R., Briskin, D.P., Wyse, R.E. (1986) Ca2+ uptake by endoplasmic reticulum from zucchini hypocotyls. Plant Physiol. 82, 47–53

    Google Scholar 

  • Ling, V., Zielinski, R.E. (1989) Cloning of cDNA sequences encoding the calcium binding protein, calmodulin from barley (Hordeum vulgare L.). Plant Physiol. 90, 714–719

    Google Scholar 

  • Malatialy, L., Greppin, H., Penel, C. (1988) Ca2+ uptake by tonoplast and plasma membrane vesicles from spinach leaves. FEBS Lett. 233, 196–200

    Google Scholar 

  • Muto, S., Miyachi, S. (1977) Properties of a protein activator of NAD kinase from plants. Plant Physiol. 59, 55–60

    Google Scholar 

  • Rasi-Caldogno, F., De Michelis, M.I., Pugliarello, M.C. (1989) Active transport of Ca2+ in membrane vesicles from pea. Evidence for a H +/Ca2+ antiport. Biochim. Biophys. Acta 693, 287–295

    Google Scholar 

  • Richmann, P.G., Klee, C.B. (1978) Interaction of 125I-labelled Ca2+-dependent regulator protein with cyclic nucleotide phosphodiesterase and its inhibitory protein. J. Biol. Chem. 253, 6323–6326

    Google Scholar 

  • Roberts, D.M., Crea, R., Malecha, M., Alvarad-Urbina, G., Chirello, R.H., Watterson, D.M. (1985) Chemical synthesis and expression of a calmodulin gene designed for site-specific mutagenesis. Biochemistry 24, 5090–5098

    Google Scholar 

  • Roberts, D.M., Harmon, A.C. (1992) Calcium modulated proteins: targets of intracellular calcium signals in higher plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43, 375–414

    Google Scholar 

  • Robinson, C., Larsson, C., Buckhout, T.J. (1988) Identification of a calmodulin stimulated (Ca2+/Mg2+)-ATPase in a plasma membrane fraction isolated from maize (it{Zea mays) leaves}. Physiol. Plant. 72, 177–184

    Google Scholar 

  • Schatzmann, H.J. (1989) The calcium pump of the surface membrane and the sarcoplasmic reticulum. Annu. Rev. Physiol. 51, 473–485

    Google Scholar 

  • Shore, G., MacLachlan, G.A. (1975) The site of cellulose synthesis. J. Cell Biol. 64, 557–571

    Google Scholar 

  • Sticher, L., Jones, R.L. (1992) α-Amylase isoforms are posttranslationally modified in the endomembrane system of the barley aleurone layer. Plant Physiol. 92, 1080–1086

    Google Scholar 

  • Tsien, R.Y., Tsien R.W. (1990) Calcium channels, stores, and oscillations. Annu. Rev. Cell Biol. 6, 715–760

    Google Scholar 

  • Villa A., Podini, P., Clegg, D.O., Pozzan, T., Medollesi, E. (1991) Intracellular Ca2+ stores in chicken Purkinje neurons: differential distribution of the low affinity, high capacity Ca2+ binding protein, calsequestrin, of Ca2+ — ATPase and of the ER lumenal protein, BiP. J. Cell Biol. 113, 779–791

    Google Scholar 

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Authors and Affiliations

  1. Department of Plant Biology, University of California, 94720, Berkeley, CA, USA

    Simon Gilroy & Russell L. Jones

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  1. Simon Gilroy
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  2. Russell L. Jones
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We thank Belen Garrido for excellent technical assistance, Eleanor Crump for help in preparation of the manuscript and Professor Daniel Roberts (Biochemistry Department, University of Tennessee, USA) for the CaM RIA kit. This work was supported by grants to R.L.J. from the National Science Foundation.

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Gilroy, S., Jones, R.L. Calmodulin stimulation of unidirectional calcium uptake by the endoplasmic reticulum of barley aleurone. Planta 190, 289–296 (1993). https://doi.org/10.1007/BF00196956

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