Skip to main content
SpringerLink
Log in

Molecular cloning and characterization of a gibberellin-inducible, putative α-glucosidase gene from barley

  • Research article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

A putative α-glucosidase clone has been isolated from a cDNA library constructed from mRNA of barley aleurones treated with gibberellin A3 (GA). The clone is 2752 bp in length and has an uninterrupted open reading frame encoding a polypeptide of 877 amino acids. A 680 amino acid region is 43% identical to human lysosomal α-glucosidase and other glycosyl hydrolases. In isolated aleurones, the levels of the corresponding mRNA increase strongly after the application of GA, similar to the pattern exhibited by low-pI α-amylase mRNA. High levels are also observed in the aleurone and scutellum after germination, while low levels are found in developing seeds. The genome contains a single form of this α-glucosidase gene and two additional sequences that may be related genes or pseudogenes.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Belanger FC, Brodl MR, Ho T-H D: Heat shock causes destabilization of specific mRNAs and destruction of endoplasmic reticulum in barley aleurone cells. Plant Physiol 83: 1354–1358 (1986).

    Google Scholar 

  2. Benton WD, Davis RW: Screening λgt recombinant clones by hybridization to single plaques in situ. Science 196: 180–182 (1977).

    Google Scholar 

  3. Blish MT, Sandstedt RM, Mecham DK: Action of wheat amylases on raw wheat starch. Cereal Chem 14: 605–628 (1937).

    Google Scholar 

  4. Chantret I, Lacasa M, Chevalier G, Ruf J, Islam I, Mantei N, Edwards Y, Swallow D, Rousset M: Sequence of the complete cDNA and the 5′ structure of the human sucrase-isomaltase gene. Possible homology with a yeast glucoamylase. Biochem J 285: 915–923 (1992).

    Google Scholar 

  5. Chrispeels MJ, Varner JE: Gibberellic acid-enhanced synthesis and release of α-amylase and ribonuclease by isolated aleurone layers. Plant Physiol 42: 398–406 (1967).

    Google Scholar 

  6. Clutterbuck VJ, Briggs DE: Enzyme formation and release by isolated barley aleurone layers. Phytochemistry 12: 537–546 (1973).

    Google Scholar 

  7. Coutinho PM, Reilly PJ: Structure-function relationships in the catalytic and starch binding domains of glucoamylase. Protein Engng 7: 393–400 (1994).

    Google Scholar 

  8. Dohmen RJ, Strasser AWM, Dahlems UM, Hollenberg CP: Cloning of the Schwanniomyces occidentalis glucoamylase gene (GAM1) and its expression in Saccharomyces cerevisiae. Gene 95: 111–121 (1990).

    Google Scholar 

  9. Dunn G: A model for starch breakdown in higher plants. Phytochemistry 13: 1341–1346 (1974).

    Google Scholar 

  10. Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specificity. Anal Biochem 132: 6–13 (1983).

    Google Scholar 

  11. Fincher GB, Lock PA, Morgan MM, Lingelbach K, Wettenhall REH, Mercer JFB, Brandt A, Thomsen KK: Primary structure of the (1–3, 1–4)-β-D-glucan 4-glucohydrolase from barley aleurone. Proc Natl Acad Sci USA 83: 2081–2085 (1986).

    Google Scholar 

  12. Geber A, Williamson PR, Rex JH, Sweeney EC, Bennett JE: Cloning and characterization of a Candida albicans maltase gene involved in sucrose utilization. J Bact 174: 6992–6996 (1992).

    Google Scholar 

  13. Genetics Computer Group, University of Wisconsin, Sequence Analysis Software, Wisconsin Package, Version 8. 1 (1994).

  14. Green F, Edwards Y, Hauri H-P, Povey S, Ho MW, Pinto M, Swallow D: Isolation of a cDNA probe for a human jejunal brush-border hydrolase, sucrase-isomaltase, and assignment of the gene locus to chromosome 3. Gene 57: 101–110 (1987).

    Google Scholar 

  15. Hardie DG: Control of carbohydrase formation by gibberellic acid in barley endosperms. Phytochemistry 14: 1719–1722 (1975).

    Google Scholar 

  16. Hata Y, Tsuchiya K, Kitamoto K, Gomi K, Kumagai C, Tamura G, Hara S: Nucleotide sequence and expression of the glucoamylase-encoding gene (glaA) from Aspergillus oryzae. Gene 108: 145–150 (1991).

    Google Scholar 

  17. Henrissat B, Bairoch A: New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 293: 781–788 (1993).

    Google Scholar 

  18. Hermans MMP, Kroos MA, van Beeumen J, Oostra BA, Reuser AJJ: Human lysosomal α-glucosidase. Characterization of the catalytic site. J Biol Chem 266: 13507–13512 (1991).

    Google Scholar 

  19. Hoefsloot LH, Hoogeveen-Westerveld M, Kroos MA, Van Beeuman J, Reuser AJJ, Oostra BA: Primary structure and processing of lysosomal α-glucosidase; homology with the intestinal sucrase-isomaltase complex. EMBO J 7: 1697–1704 (1988).

    Google Scholar 

  20. Hong SH, Marmur J: Primary structure of the maltase gene of the Mal 6 locus of Saccharomyces carlsbergensis. Gene 41: 75–84 (1984).

    Google Scholar 

  21. Hostinova E, Balanova J, Gasperik J: The nucleotide sequence of the glucoamylase gene GLA1 from Saccharomypsis fibuligera KZ. FEMS Microbiol Lett 83: 103–108 (1991).

    Google Scholar 

  22. Hunziker W, Spiess M, Semenza G, Lodish HF: The sucrase-isomaltase complex: Primary structure, membrane-orientation, and evolution of a stalked, intrinsic brush border protein. Cell 46: 227–234 (1986).

    Google Scholar 

  23. James AA, Blackmer K, Racioppi JV: A salivary gland-specific maltase-like gene of the vector mosquito, Aedes aegypti. Gene 75: 73–83 (1989).

    Google Scholar 

  24. Janse BJH, Steyn AJC, Pretorius IS: Regional sequence homologies in starch-degrading enzymes. Curr Genet 24: 400–407 (1993).

    Google Scholar 

  25. Jespersen HM, MacGregor EA, Henrissat B, Sierks MR, Svensson B: Starch- and glycogen-debranching and branching enzymes: Prediction of structural features of the catalytic (β/α)8-barrel domain and evolutionary relationship to other amylolytic enzymes. J Prot Chem 12: 791–805 (1993).

    Google Scholar 

  26. Jorgensen BB, Jorgensen OB: Barley malt α-glucosidase with isomaltase activity. Acta Chem Scand 17: 1765–1770 (1963).

    Google Scholar 

  27. Jorgensen OB: Barley malt α-glucosidase. VI. Localization and development during barley germination. Acta Chem Scand 19: 1014–1015 (1965).

    Google Scholar 

  28. Kinsella BT, Hogan S, Larkin A, Cantwell BA: Primary structure and processing of the Candida tsukubaensis α-glucosidase. Homology with the rabbit intestinal sucrase-isomaltase complex and human lysosomal α-glucosidase. Eur J Biochem 202: 657–664 (1991).

    Google Scholar 

  29. Kislev N, Rubenstein I: Utility of ethidium bromide in the extraction from whole plants of high molecular weight maize DNA. Plant Physiol 66: 1140–1143 (1980).

    Google Scholar 

  30. Konishi Y, Okamoto A, Takahashi J, Aitani M, Nakatani N: Effects of Bay m 1099, an α-glucosidase inhibitor, on starch metabolism in germinating wheat seeds. Biosci Biotechnol Biochem 58: 135–139 (1994).

    Google Scholar 

  31. Koehler SM, Ho T-H D: Hormonal regulation, processing and secretion of cysteine proteinases in barley aleurone layers. Plant Cell 2: 769–783 (1990).

    Google Scholar 

  32. Kopetzki E, Buckel P, Schumacher G: Cloning and characterization of Baker's yeast α-glucosidase: over-expression in a yeast strain devoid of vacuolar proteinases. Yeast 5: 11–24 (1989).

    Google Scholar 

  33. Kozak M: Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44: 283–292 (1986).

    Google Scholar 

  34. Kreis M, Williamson M, Buxton B, Pywell J, Hejgaard J, Svendsen I: Primary structure and differential expression of β-amylase in normal and mutant barleys. Eur J Biochem 169: 517–525 (1987).

    Google Scholar 

  35. MacGregor AW, Lenoir C: Studies on α-glucosidase in barley malt. J Inst Brew 93: 334–337 (1987).

    Google Scholar 

  36. Martiniuk F, Mehler M, Tzall S, Meredith G, Hirschhorn R: Sequence of the cDNA and 5′-flanking region for human acid α-glucosidase, detection of an intron in the 5′ untranslated leader sequence, definition of 18-bp polymorphisms, and differences with previous cDNA and amino acid sequences. DNA Cell Biol 9: 85–94 (1990).

    Google Scholar 

  37. Naim HY, Niermann T, Kleinhans U, Hollenberg CP, Strasser AWM: Striking structural and functional similarities suggest that intestinal sucrase-isomaltase, human lysosomal α-glucosidase and Schwanniomyces occidentalis glucoamylase are derived from a common ancestral gene. FEBS Lett 294: 109–112 (1991).

    Google Scholar 

  38. Nakao M, Nakayama T, Kakudo A, Inohara M, Harada M, Omura F, Shibano Y: Structure and expression of a gene coding for thermostable α-glucosidase with a broad substrate specificity from Bacillus sp. SAM1606. Eur J Biochem 220: 293–300 (1994).

    Google Scholar 

  39. Rave N, Crkvenjakov R, Boedtker H: Identification of procollagen mRNAs transferred to diazobenzyloxymethyl paper from formaldehyde agarose gels. Nucl Acids Res 6: 3559–3567 (1979).

    Google Scholar 

  40. Rick W, Stegbauer HP: Measurement of reducing groups. In: Bergmeyer HU (ed) Methods of Enzymatic Analysis, 2nd ed., vol 2, pp. 885–889. Academic Press, New York (1974).

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  43. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed., pp. 1.42–1.46, 9.38–9.40. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  44. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    Google Scholar 

  45. Sierks MR, Ford C, Reilly PJ, Svensson B: Functional roles and subsite locations of Leu 177, Trp 178 and Asn 182 of Aspergillus awamori glucoamylase determined by sitedirected mutagenesis. Protein Engng 6: 75–79 (1993).

    Google Scholar 

  46. Skadsen RW: Aleurones from a barley with low α-amylase activity become highly responsive to gibberellin when detached from the starchy endosperm. Plant Physiol 102: 195–203 (1993).

    Google Scholar 

  47. Skadsen RW, Tibbot BK: Temporal expression patterns of alpha-amylase isozymal genes in polysomal and total RNAs of germinating barleys. J Cereal Sci 19: 199–208 (1994).

    Google Scholar 

  48. Snyder M, Davidson N: Two gene families clustered in a small region of the Drosophila genome. J Mol Biol 166: 101–118 (1983).

    Google Scholar 

  49. Stark JR, Yin XS: Evidence for the presence of maltase and α-glucosidase isozymes in barley. J Inst Brew 93: 108–112 (1987).

    Google Scholar 

  50. Sun Z, Henson CA: A quantitative assessment of the importance of barley seed α-amylase, β-amylase, debranching enzyme, and α-glucosidase in starch degradation. Arch Biochem Biophys 284: 298–305 (1991).

    Google Scholar 

  51. Sun Z, Henson CA: Degradation of native starch granules by barley α-glucosidases. Plant Physiol 94: 320–327 (1990).

    Google Scholar 

  52. Svensson B: Regional distant sequence homology between amylases, α-glucosidases and transglucanosylases. FEBS Lett 230: 72–76 (1988).

    Google Scholar 

  53. Tanaka Y, Ashikari T, Nakamura N, Kiuchi N, Shibano Y, Amachi T, Yoshizumo H: Comparison of amino acid sequence of three glucoamylases and their structure-function relationships. Agric Biol Chem 50: 965–969 (1986).

    Google Scholar 

  54. Tapio S, Yeh F, Shuman HA, Boos W: The malZ gene of Escherichia coli, a member of the maltose regulon, encodes a maltodextrin glucosidase. J Biol Chem 266: 19450–19458 (1991).

    Google Scholar 

  55. Thomas PS: Hybridization of denatured RNA transferred or dotted to nitrocellulose paper. Meth Enzymol 100: 255–266 (1983).

    Google Scholar 

  56. Watson TG, Novellie L: Extraction of Sorghum vulgare and Hordeum vulgare α-glucosidase. Phytochemistry 13: 1037–1041 (1974).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Agronomy, University of Wisconsin, 53706, Madison, WI, USA

    Brian K. Tibbot & Ronald W. Skadsen

  2. Cereal Crops Research Unit, USDA, Agricultural Research Service, 53705, Madison, WI, USA

    Brian K. Tibbot & Ronald W. Skadsen

Authors
  1. Brian K. Tibbot
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ronald W. Skadsen
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tibbot, B.K., Skadsen, R.W. Molecular cloning and characterization of a gibberellin-inducible, putative α-glucosidase gene from barley. Plant Mol Biol 30, 229–241 (1996). https://doi.org/10.1007/BF00020110

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00020110

Key words

Search

Navigation