Skip to main content

Regulation of the expression of α-amylase gene by sodium butyrate

Plant Molecular Biology volume 5, pages 269–279 (1985)Cite this article

Summary

Sodium butyrate exerts a pronounced inhibition on the gibberellic acid-induced synthesis and secretion of α-amylase by aleurone cells of barley seeds. This inhibition, which is reversible and non-competitive with cespect to gibberellic acid, is concentration dependent, with virtually total inhibition being accomplished between 4 and 5 mM sodium butyrate. The pattern of inhibition of α-amylase formation correlates well with a decrease in the accumulation of its messenger RNA. The addition of butyrate 12 h after the addition of gibberellic acid to half-seeds, has no effect on the formation and secretion of α-amylase. It has been shown in earlier studies that the synthesis of α-amylase mRNAs takes about 12 h for completion. The conclusion that butyrate interferes with some step in the transcriptional process is supported by a decrease observed in the RNAs that hybridize with a cloned α-amylase cDNA. The results of in vitro translation confirm the inhibition of the formation of several translatable mRNAs. Further, immunological probing detected only trace amounts of α-amylase proteins in the secretion of butyrate-treated seeds. Translation of functional mRNAs, post-translational modifications and the secretion α-amylase are not affected by sodium butyrate. It is concluded that butyrate selectively inhibits the transcription of several genes that are under the influence of gibberellic acid. This report is the first one documenting the inhibitory effect of sodium butyrate on a hormone-induced synthesis and accumulation of mRNAs in a plant system.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Abbreviations

GA:

gibberellic acid

NaOBut:

sodium butyrate

DBM:

diazobenzyloxymethyl

References

  1. Abdinejad A, Fisher A, Kumar S: Production and utilization of butyryl-CoA by fatty acid synthetase from mammalian tissue. Arch Biochem Biophys 208:135–145, 1981.

    Google Scholar 

  2. Akazawa T, Miyata S: Biosynthesis and secretion of α-amylase and other hydrolases in germinating cereal seeds. Essays in Biochemistry 18:40–78, (1984).

    Google Scholar 

  3. Alwine JC, Kemp DJ, Stark GR: Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl paper. Proc Natl Acad Sci USA 74:5350–5354, 1977.

    Google Scholar 

  4. Alwine JC, Kemp DJ, Parker GA, Reiser J, Renart J, Stark GR, Wahl GM: Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods in Enzymology 68: 220–242, 1980.

    Google Scholar 

  5. Bell PA, Jones CN: Cytotoxic effects of butyrate and other ‘differentiation inducers’ on immature lymphoid cells. Biochem Biophys Res Commun 104:1202–1208, 1982.

    Google Scholar 

  6. Bernal-Lugo I, Beachy RN, Varner JE: The response of barley aleurone layers to gibberellic acid includes the transcription of new sequences. Biochem Biophys Res Commun 102:617–623, 1981.

    Google Scholar 

  7. Boffa LC, Gruss RJ, Allfrey VG: Manifold effects of sodium butyrate on nuclear function. Selective and reversible inhibition of phosphorylation. J Biol Chem 256:9612–9621, 1981.

    Google Scholar 

  8. Brown AHD, Jacobsen JV: Genetic basis and natural variation of α-amylase isozymes in barley. Genet Res 40:315–324, 1982.

    Google Scholar 

  9. Buller DC, Parker W, Reid JSG: Short chain fatty acids are inhibitors of GA-induced amylolysis in barley endosperm. Nature 260:169–170, 1976.

    Google Scholar 

  10. Callis J, Ho T-h.D: Multiple molecular forms of the gibberellin-induced α-amylase from the aleurone layers of barley seeds. Arch Biochem Biophys: 224–234, 1983.

  11. Candiodo EP, Reeves R, Davie JR: Sodium butyrate inhibits histone deacetylation in cultured cells. Cell 14:105–113, 1978.

    Google Scholar 

  12. Chandler PM, Zwar JA, Jacobsen JV, Higgins TJV, Inglis AS: The effects of gibberellic acid and abscisic acid on α-amylase mRNA levels in barley aleurone layers studies using an α-amylase cDNA clone. Plant Molecular Biol 3:407–418, 1984.

    Google Scholar 

  13. Chandra GR, Muthukrishnan S, Maxwell ES: Hormonal regulation of genome activity in higher plants, ACS, Symposium Series, No 111, Plant Growth Substances, 245–261, 1979.

  14. Davie JR, Saunders CA: Chemical composition of nucleosomes among domains of calf thymus chromatin differing in micrococcal nuclease accessibility and solubility properties. J Biol Chem 256:12574–12580, 1981.

    Google Scholar 

  15. Filner P, Varner JE: A test for de novo synthesis of enzymes: density labeling with H2O18 of barley α-amylase induced by gibberellic acid. Proc Natl Acad Sci USA 58: 1520–1526, 1967.

    Google Scholar 

  16. Ghosh NK, Cox RP: Induction of human follicle stimulating hormone in HeLa cells by sodium butyrate. Nature 267: 435–437, 1977.

    Google Scholar 

  17. Herz F, Halwer M: Synergistic induction of alkaline phosphatase in colonic carcinoma cells by sodium butyrate and hyperiesmolality. Biochim Biophys Acta 718:220–223, 1982.

    Google Scholar 

  18. Higgins TJV, Zwar JA, Jacobsen JV: Gibberellic acid enhances the level of translatable mRNA for α-amylase in barley aleurone layers. Nature 260:166–169, 1976.

    Google Scholar 

  19. Higgins TJV, Zwar JA, Jacobsen JV: Gibberellic acid and abscisic acid modulate protein synthesis and mRNA levels in barley aleurone layers. Plant Molec Biol 1:191–215, 1982.

    Google Scholar 

  20. Ho T-h.D, Hormonal and genetic regulation of α-amylase synthesis in barley aleurone cells. In: Lever CJ (ed) Genome organization and expression in plants. Plenum, New York, London, 1980, pp 147–157.

    Google Scholar 

  21. Ho T-h.D, Varner JE: Density labeling of proteins with 13C-labeled amino acids. No accumulation of an inactive α-amylase precursor in barley aleurone cells. Arch Biochem Biophys 187:441–446, 1978.

    Google Scholar 

  22. Huang JK, Swegle M, Dandekar AM, Muthukrishnan S: Expression and regulation of α-amylase gene family in barley aleurone. J Mol Appl Genet, in press.

  23. Jacobsen JV: Regulation of ribonucleic acid metabolism by plant hormones. Ann Rev Plant Physiol 28:537–564, 1977.

    Google Scholar 

  24. Jacobsen JV, Regulation of protein synthesis in aleurone cells by gibberellin and abscisic acid. In: Crozier A (ed) The biochemistry and physiology of gibberellins, Vol 2. Praeger, New York, 1984.

    Google Scholar 

  25. Jacobsen JV, Higgins TJV: Characterization of the α-amylase synthesized by aleurone layers of Himalaya barley in response to GA3. Plant Physiol 70:1647–1653, 1982.

    Google Scholar 

  26. Jones RL, Varner JE: The bioassay of gibberellins. Planta (Ber.) 72:155–161, 1967.

    Google Scholar 

  27. Kitzis A, Tichonicky L, Defer N, Kruh J: Effect of sodium butyrate on chromatin structure. Biochem Biophys Res Commun 93:883–841, 1980.

    Google Scholar 

  28. Koshihara Y, Senshu T, Kawamura M, Murota S: Sodium n-butyrate induces prostaglandin synthetase activity in mastocytoma P-815 cells. Biochim Biophys Acta 617:536–539, 1980.

    Google Scholar 

  29. Kruh J: Effects of sodium butyrate, a new pharmacological agent, on cells in culture. Mol Cellular Biochem 42:65–82, 1981.

    Google Scholar 

  30. Leder A, Leder P: Butyric acid, a protein inducer of erythroid differentiation in cultured erythroleukemic cells. Cell 5: 319–322, 1975.

    Google Scholar 

  31. Martel P, Houdebine LM, Teyssot B: Effect of sodium butyrate on the stimulation of casein gene expression by prolactin. FEBS Letters 154:55–59, 1983.

    Google Scholar 

  32. McKnight GS, Hager L, Palmiter R: Butyrate and related inhibitors of histone deacetylation block the induction of egg white genes by steroid hormones. Cell 22:469–477, 1980.

    Google Scholar 

  33. McMaster GK, Carmichael GG: Analysis of single and double stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci USA 74:4835–4838, 1977.

    Google Scholar 

  34. Meredith P: Do fatty acids inhibit gibberellin-induced amylolysis? Nature 270:193, 1977.

    Google Scholar 

  35. Morser J, Meager A, Colman A: Enhancement of interferon mRNA levels in butyric acid-treated namalwa cells. FEBS Letters 112:203–206, 1980.

    Google Scholar 

  36. Morrow JS, Weintraub BD, Rosen S: Butyrate regulates glycosylation of the glycoprotein hormone alpha subunit secreted by ‘glucose-starved’ human liver cells. Biochem Biophys Res Commun 112:115–125, 1983.

    Google Scholar 

  37. Mozer TJ: Control of protein synthesis in barley aleurone layers by the plant hormones gibberellic acid and abscisic acid. Cell 20:479–485, 1980.

    Google Scholar 

  38. Muthukrishnan S, Chandra GR, Maxwell ES: Hormone-induced increase in levels of functional mRNA and α-amylase mRNA in barley aleurones. Proc Natl Acad Sci USA 76:6181–6185, 1979.

    Google Scholar 

  39. Muthukrishnan S, Chandra GR: Expression of α-amylase genes in barley aleurone cells. In: Owens LD (ed) Genetic Eng Appl to Agric (Beltsville Symp 7). Rowman and Allanheld, Totowa.

  40. Muthukrishnan S, Chandra GR, Maxwell ES: Hormonal control of α-amylase gene expression in barley studies using a cloned cDNA probe. J Biol Chem 258:2370–2375, 1983.

    Google Scholar 

  41. Muthukrishnan S, Chandra GR, Albaugh GP: Modulation by abscisic acid and S-2 aminoethyl-L-cysteine of α-amylase mRNA in barley aleurone cells. Plant Mol Biol 2:249–258, 1983.

    Google Scholar 

  42. Muthukrishnan S, Gill BS, Swegle M, Chandra GR: Structural genes for α-amylases are located on barley chromosomes 1 and 6. J Biol Chem 259:3637–3639, 1984.

    Google Scholar 

  43. Nelson D, Covault J, Perry WM, Chalkley R: Segregation of rapidly acetylated histones into a chromatin fraction released from intact nuclei by the action of micrococcal nuclease. Nucleic Acids Res 8:1745–1763, 1980.

    Google Scholar 

  44. Plesko MM, Hargroves JL, Granner DK, Chalkley R: Inhibition by sodium butyrate of enzyme induction by gluco-corticoids and dibutyryl cyclic AMP. A role for the rapid form of histone acetylation. J Biol Chem 258:13738–13744, 1983.

    Google Scholar 

  45. Prasad KN: Butyric acid: A small fatty acid with diverse biological functions. Life Sci 27:1351–1358, 1980.

    Google Scholar 

  46. Reczek RP, Weissman D, Huvos P, Fasman GD: Sodium butyrate induced structural changes in HeLa cell chromatin. Biochem 21:993–1002, 1982.

    Google Scholar 

  47. Reeves R, Cserjesi P: Sodium butyrate induces new gene expression in Friend erythroleukemic cells. J Biol Chem 254:4283–4290, 1979.

    Google Scholar 

  48. Rigby PWJ, Dieckmann M, Rhodes C, Berg P: Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251.

  49. Rodaway SJ: Composition of α-amylase secreted by aleurone layers of grain of himalaya barley. Phytochemistry 17:385–389, 1978.

    Google Scholar 

  50. Rogers JC, Milliman C: Isolation and sequence analysis of a barley α-amylase cDNA clone. J Biol Chem 258:8169–8178, 1983.

    Google Scholar 

  51. Rogers JC, Milliman C: Coordinate increase in major transcripts from the high pl α-amylase multigene family in barley alcurone cells stimulated with gibberellic acid. J Biol Chem 259:12234–12240, 1984.

    Google Scholar 

  52. Sealy L, Chalkley R: The effect of sodium butyrate on histone modification. Cell 14:115–121, 1978.

    Google Scholar 

  53. Thomas P: Hybridization of denatured RNA and small DNA fragments transferred to mitrocellulose. Proc Natl Acad Sci USA 77:5201–5205, 1980.

    Google Scholar 

  54. Varner JE, Chandra GR: Hormonal control of enzyme synthesis in barley endosperm. Proc Natl Acad Sci 52:100–106, 1964.

    Google Scholar 

  55. Waymire C, Weiner N, Prasad KN: Regulation of tyrosine hydroxylase activity in cultured mouse neurolpastorna cells: Elevation induced by analogs of adenesine 3′:5′-cyclic monophosphate. Proc Natl Acad Sci USA 69:2241–2245, 1972.

    Google Scholar 

  56. Yomo H, Varner JE: Hormonal control of a secretory tissue. Curr Topics Develop Biol 6:111–144, 1974.

    Google Scholar 

Download references

Author information

Affiliations

  1. Department of Chemistry, Georgetown University, 20057, Washington, DC, U.S.A.

    Soma Kumar

  2. Seed Research Laboratory, Beltsville Agricultural Research Center-West, 20705, Beltsville, MD, U.S.A.

    G. Ram Chandra

  3. Department of Botany, University of Maryland, 20742, College Park, MD, U.S.A.

    G. P. Albaugh

  4. Department of Biochemistry, Kansas State University, 66506, Manhattan, KS, U.S.A.

    S. Muthukrishnan

Authors
  1. Soma Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Ram Chandra
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. P. Albaugh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S. Muthukrishnan
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kumar, S., Chandra, G.R., Albaugh, G.P. et al. Regulation of the expression of α-amylase gene by sodium butyrate. Plant Mol Biol 5, 269–279 (1985). https://doi.org/10.1007/BF00020624

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00020624

Key words

  • α-amylase
  • barley
  • sodium butyrate
  • gene
  • gibberellic acid
  • mRNA