Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

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

  • 58 Accesses

  • 31 Citations

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.

This is a preview of subscription content, log in to check access.

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

  1. 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

  2. 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

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

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

  5. 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

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

  7. 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

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

  9. 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

  10. 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

  11. 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

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

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

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

  15. 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

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

  17. 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

  18. 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

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

  20. 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

  21. 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

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

  23. 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

  24. 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

  25. 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

  26. 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

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

  28. 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

  29. 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

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

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

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

  33. 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

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

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

  36. 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

  37. 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

  38. 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

  39. 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

  40. 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

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

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

  43. 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

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

  45. 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

Download references

Author information

Correspondence to Russell L. Jones.

Additional information

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.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

Key words

  • Aleurone
  • Calcium uptake
  • Calmodulin
  • Endoplasmic reticulum
  • Gibberellic acid
  • Hordeum
  • (Ca2+ uptake)