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Selective expression of a probable amylase/protease inhibitor in barley aleurone cells: Comparison to the barley amylase/subtilisin inhibitor

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Abstract

We have cloned and sequenced a 650-nucleotide cDNA from barley (Hordeum vulgare L.) aleurone layers encoding a protein that is closely related to a known α-amylase inhibitor from Indian finger millet (Eleusine coracana Gaertn.), and that has homologies to certain plant trypsin inhibitors. mRNA for this probable amylase/protease inhibitor (PAPI) is expressed primarily in aleurone tissue during late development of the grain, as compared to that for the amylase/subtilisin inhibitor, which is expressed in endosperm during the peak of storage-protein synthesis. PAPI mRNA is present at high levels in aleurone tissue of desiccated, mature grain, and in incubated aleurone layers prepared from rehydrated mature seeds. Its expression in those layers is not affected by either abscisic acid or gibberellic acid, hormones that, respectively, increase and decrease the abundance of mRNA for the amylase/subtilisin inhibitor. PAPI mRNA is almost as abundant in gibberellic acid-treated aleurone layers as that for α-amylase, and PAPI protein is synthesized in that tissue at levels that are comparable to α-amylase. PAPI protein is secreted from aleurone layers into the incubation medium.

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Abbreviations

ABA:

abscisic acid

ASI:

barley amylase/subtilisin inhibitor

bp:

nucleotide base pairs

Da:

dalton

dpa:

days post anthesis

GA3 :

gibberellic acid

PAPI:

probable amylase/protease inhibitor

poly(A)RNA:

polyadenylated RNA

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

References

  • Anderson, D.J. Blobel, G. (1983) Immunoprecipitation of proteins from cell-free translations. Methods Enzymol.96, 111–120

    PubMed  Google Scholar 

  • Boisen, S., Andersen, C.Y. Hejgaard, J. (1981) Inhibitors of chymotrypsin and microbial serine proteases in barley grains. Physiol. Plant.52, 167–176

    Google Scholar 

  • Bollum, F.J. (1975) Antibody to terminal deoxynucleotidyl transferase. Proc. Natl. Acad. Sci USA72, 4119–4122

    PubMed  Google Scholar 

  • Campos, F.A.P., Richardson, M. (1983) The complete amino acid sequence of the bifunctional α-amylase inhibitor from seeds of ragi (Indian finger millet,Eleusine coracana Gaertn. FEBS. Lett.152, 300–304

    Google Scholar 

  • Campos, F.A.P., Richardson, M. (1984) The complete amino acid sequence of the α-amylase inhibitor I-2 from seeds of ragi (Indian finger millet,Eleusine coracana Gaertn.). FEBS Lett.167, 221–225

    Google Scholar 

  • Croy, R.R., Lycett, G., Gatehouse, J.A., Yarwood, J.N., Boulter, D. (1982) Cloning and analysis of cDNAs encoding plant storage protein precursors. Nature295, 76–78

    Google Scholar 

  • Dayhoff, M.O. Barker, W.C., Hunt, L.T. (1983) Establishing homologies in protein sequences. Methods Enzymol.91, 524–545

    PubMed  Google Scholar 

  • Giese, H., Hejgaard, J. (1984) Synthesis of salt-soluble proteins in barley. Pulse-labeling study of grain filling in liquid-cultured detached spikes. Planta161, 172–177

    Google Scholar 

  • Giese, H., Hopp, H.E. (1984) Influence of nitrogen nutrition on the amount of hordein, protein Z, and β-amylase messenger RNA in developing endosperms of barley. Carlsberg Res. Commun.49, 365–383

    Google Scholar 

  • Hejgaard, J., Rasmussen, S.K., Brandt, A., Svendsen, I. (1985) Sequence homology between barley endosperm protein Z and protease inhibitors of the α1-antitrypsin family. FEBS Lett.180, 89–94

    Google Scholar 

  • Hejgaard, J., Svendsen, I., Mundy, J. (1983) Barley α-amylase/subtilisin inhibitor. II. N-terminal amino acid sequence and homology with inhibitors of the soybean trypsin inhibitor (Kunitz) family. Carlsberg Res. Commun.48, 91–94

    Google Scholar 

  • Higgins, T.J.V., Jacobsen, J.V., Zwar, J.A. (1982) Gibberellic acid and abscisic acid modulate protein synthesis and mRNA levels in barley aleurone layers. Plant Mol. Biol.1, 191–215

    Google Scholar 

  • Ho, D.T-H., Varner, J.E. (1976) Response of barley aleurone layers to abscisic acid. Plant Physiol.57, 175–178

    Google Scholar 

  • Jacobsen, J.V. (1984) Regulation of protein synthesis in aleurone cells by gibberellins and abscisic acid. In: The biochemistry and physiology of gibberellins, vol. 2, pp. 159–187, Crozier, A., ed. Praeger, New York

    Google Scholar 

  • Jacobsen, J.V., Beach, L.R. (1985) Control of transcription of α-amylase and rRNA genes in barley aleurone protoplasts by gibberellin and abscisic acid. Nature316, 275–277

    Google Scholar 

  • Jensen, S.A., Munck, L., Martens, H. (1982) The botanical constituents of wheat and wheat milling fractions. I. Quantification by autofluorescence. Cereal Chem.59, 477–484

    Google Scholar 

  • Jonassen, I. (1980) Characteristics of hiproly barley II. Quantification of two proteins contributing to its high lysine content. Carlsberg Res. Commun.45, 59–68

    Google Scholar 

  • Jonassen, I., Ingversen, J., Brandt, A. (1981) Synthesis of SP II albumin, β-amylase and chymotrypsin inhibitor Cl-1 on polysomes from the endoplasmic reticulum of barley endosperm. Carlsberg Res. Commun.46, 175–181

    Google Scholar 

  • Joubert, F.J., Kruger, H., Towhshend, G.S., Botes, D.P. (1979) Purification, some properties and the complete primary structures of two protease inhibitors (DE-3 and DE-4) fromMacrotyloma axillare seed. Eur. J. Biochem.97, 85–91

    PubMed  Google Scholar 

  • Laskowski, M., Kato, I. (1980) Protein inhibitors of proteinases. Annu. Rev. Biochem.49, 593–696

    PubMed  Google Scholar 

  • Mathews, J.A., Miflin, B.J.(1980) In-vitro synthesis of barley storage proteins. Planta149, 262–268

    Google Scholar 

  • Maxam, M.A., Gilbert, W. (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol.65, 497–560

    PubMed  Google Scholar 

  • Mozer, T.J. (1980) Control of protein synthesis in barley aleurone layers by the plant hormones gibberellic acid and abscisic acid. Cell20, 479–485

    PubMed  Google Scholar 

  • Mundy, J. (1984) Hormonal regulation of α-amylase inhibitor synthesis in germinating barley. Carlsberg Res. Commun.49, 439–444

    Google Scholar 

  • Mundy, J., Brandt, A., Fincher, G. (1985) Messenger RNAs from the scutellum and aleurone of germinating barley encode (1-3, 1-4)-β-D-glucanase, α-amylase, and carboxypeptidase. Plant Physiol.79, 867–871

    Google Scholar 

  • Mundy, J., Hejgaard, J., Hansen, A., Hallgren, L., Jorgensen, K.G., Munck, L. (1986) Differential synthesis in vitro of barley aleurone and starchy endosperm proteins. Plant Physiol. (in press)

  • Mundy, J., Hejgaard, J., Svendsen, I. (1984) Characterization of a bifunctional wheat inhibitor of endogenous α-amylase and subtilisin. FEBS Lett.167, 210–214

    Google Scholar 

  • Mundy, J., Svendsen, I., Hejgaard, J. (1983) Barley α-amylase/subtilisin inhibitor. I. Isolation and characterization. Carlsberg Res. Commun.48, 81–90

    Google Scholar 

  • Odani, S., Koide, T., Ono, T. (1983) The complete amino acid sequence of barley trypsin inhibitor. J. Biol. Chem.258, 7998–8003

    PubMed  Google Scholar 

  • Ponz, F., Paz-Ares, J., Hernandez-Lucas, C., Carbonero, P., Garcia-Olmedo, F. (1983) Synthesis and processing of thionin precursors in developing endosperm from barley (Hordeum vulgare L.). EMBO J.2, 1035–1040

    Google Scholar 

  • Rasmussen, S.K., Hopp, H.E., Brandt, A. (1983) Nucleotide sequences of cDNA clones for B1 hordein polypeptides. Carlsberg Res. Commun.48, 187–199

    Google Scholar 

  • Richardson, M. (1979) The complete amino acid sequence and the trypsin reactive (inhibitory) site of the major proteinase inhibitor from the fruits of aubergine (Solanum melongena L.) FEBS Lett.104, 322–326

    PubMed  Google Scholar 

  • Rogers, J.C. (1985) Two barley α-amylase gene families are regulated differently in aleurone cells. J. Biol. Chem.260, 3731–3738

    PubMed  Google Scholar 

  • Rogers J.C., Dean, D., Heck, G.R. (1985) Aleurain: a barley thiol protease closely related to mammalian cathepsin H. Proc. Natl. Acad. Sci. USA82, 6512–6516

    PubMed  Google Scholar 

  • Rogers, J.C., Milliman, C. (1983) Isolation and sequence analysis of a barley α-amylase cDNA clone. J. Biol. Chem.258, 8169–8174

    PubMed  Google Scholar 

  • Shewry, P.R., Lafiandra, D., Salcedo, G., Aragoncillo, C., Garcia-Olmedo, F., Lew, E.J.-L., Dietler, M.D. Kasarda, D.D. (1984) N-terminal amino acid sequences of chloroform/methanol-soluble proteins and albumins from endosperms of wheat, barley and related species. FEBS Lett.175, 359–363

    Google Scholar 

  • Shivaraj, B., Pattabiraman, T.N. (1981) Natural plant enzyme inhibitors: Characterization of an unusual α-amylase/trypsin inhibitor from ragi (Eleusine coracana Geartn). Biochem. J.193, 29–36

    PubMed  Google Scholar 

  • Svensson, B., Asano, K., Jonassen, I., Poulsen, F., Mundy, J., Svendsen, I. (1986) Isolation and characterization of a barley seed protein with Mr 10,000 showing homology with an α-amylase inhibitor for Indian Finger Millet. Carlsberg Res. Commun. (in press)

  • Svensson, B., Mundy, J., Gibson, R.M., Svendsen, I. (1985) Partial amino acid sequences of α-amylase isozymes from barley malt. Carlsberg Res. Commun.50, 15–22

    Google Scholar 

  • Towbin, H., Staehelin, T., Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA76, 4350–5354

    PubMed  Google Scholar 

  • Weselake, R.J., MacGregor, A.W., Hill, R.D. (1983) An endogenous α-amylase inhibitor in barley kernels. Plant Physiol.72, 809–812

    Google Scholar 

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Authors and Affiliations

  1. Department of Biotechnology, Carlsberg Research Laboratory, Gamle Carlsberg Vej 10, DK-2500 Valby, Copenhagen, Denmark

    J. Mundy

  2. Division of Hematology/Oncology, Department of Internal Medicine, Washington University School of Medicine, 63110, St. Louis, MO, USA

    J. C. Rogers

  3. Department of Biology, Washington University School of Medicine, 63110, St. Louis, MO, USA

    J. C. Rogers

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  1. J. Mundy
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Mundy, J., Rogers, J.C. Selective expression of a probable amylase/protease inhibitor in barley aleurone cells: Comparison to the barley amylase/subtilisin inhibitor. Planta 169, 51–63 (1986). https://doi.org/10.1007/BF01369775

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  • DOI: https://doi.org/10.1007/BF01369775

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