Abstract
A wheat gene (A121) encoding a protein with sequence similarity to mammalian cathepsin B is regulated by gibberellic acid (GA) in aleurone layers of germinating grains. To analyse the mechanism of A121 regulation, its promoter was fused to the β-glucuronidase reporter gene (GUS) and introduced by micro-projectile bombardment into aleurone layers of oat. With 2.3 kb of promoter sequence, the GUS expression was enhanced by GA treatment. This effect was reversed by abscisic acid (ABA). This result showed for A121, like the α-amylase genes, that the regulation by GA and ABA was at the level of transcription. The GA responsiveness of the promoter was retained with as little as 276 bp of promoter sequence. Sequence comparison with a GA responsive promoter of an α-amylase gene identified the conserved element GCAACGGCAACGATGG which is required intact for full expression of both promoters. However, there was no identifiable similarity in the cathepsin-like promoter with the GA-responsive element of α-amylase promoters with the consensus sequence TAACAAA, suggesting that GA affects more than one mechanism of transcriptional control.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baulcombe DC, Barker RF, Jarvis MG: A gibberellin responsive wheat gene has homology to yeast carboxypeptidase Y. J Biol Chem 262: 13726–13735 (1987).
Baulcombe DC, Buffard D: Gibberellic-acid-regulated expression of α-amylase and six other genes in wheat aleurone layers. Planta 157: 493–501 (1983).
Baulcombe DC, Huttly AK, Martienssen RA, Barker RF, Jarvis MG: A novel wheat α-amylase gene (α-Amy3). Mol Gen Genet 209: 33–40 (1987).
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 Mol Biol 3: 407–418 (1984).
Chandler PM, Jacobsen JV: Primer extension studies on a-amylase mRNAs in barley aleurones. II. Hormonal regulation of expression. Plant Mol Biol 16: 637–645 (1991).
Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).
Fincher GB: Molecular and cellular biology associated with endosperm mobilization in germinating cereal grains. Annu Rev Plant Physiol Plant Mol Biol 40: 305–346 (1989).
Gorman CM, Moffat LF, Howard BH: Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 2: 1044–1051 (1982).
Hammerton RW, Ho T-HDC: Hormonal regulation of the development of protease and carboxypeptidase activities in barley aleurone layers. Plant Physiol 80: 692–697 (1986).
Ho T-HD, Nolan RC, Shute DE: Characterization of a gibberellin-insensitive dwarf wheat, D6899. Plant Physiol 67: 1026–1031 (1981).
Huang N, Sutliff TD, Litts JC, Rodriguez RL: Classification and characterization of the rice α-amylase multigene family. Plant Mol Biol 14: 655–658 (1990).
Huttly AK, Baulcombe DC: A wheat α-Amy2 promoter is regulated by gibberellin in transformed oat aleurone protoplasts. EMBO J 8: 1907–1913 (1989).
Huttly AK, Martienssen RA, Baulcombe DC: Sequence heterogeneity and differential expression of the α-Amy2 gene family in wheat. Mol Gen Genet 214: 232–240 (1989).
Jacobsen JV, Beach LR: Control of transcription of α-amylase and rRNA genes in barley aleurone protoplasts by gibberellin and abscisic acid. Nature 316: 275–277 (1985).
Jacobsen JV, Close TJ: Control of transient expression of chimaeric genes by gibberellic acid and abscisic acid in protoplasts prepared from mature barley aleurone layers. Plant Mol Biol 16: 713–724 (1991).
Klein TM, Gradziel T, Fromm ME, Sanford JC: Factors in influencing gene delivery into Zea mays cells by high-velocity microprojectiles. Bio/technology 6: 559–563 (1988).
Koehler SM, Ho T-HD: Hormone regulation, processing, and secretion of cysteine proteases in barley aleurone layers. Plant Cell 2: 769–7833 (1990).
Lanahan MB, Ho T-HD, Rogers SW, Rogers JC: A gibberellin response complex in cereal α-amylase gene promoters. Plant Cell, in press.
Lazarus CM, Baulcombe DC, Martienssen RA: α-amylase genes of wheat are two multigene families which are differentially expressed. Plant Mol Biol 5: 13–24 (1985).
Maas C, Laufs J, Grant S, Korfhage C, Werr W: The combination of a novel stimulatory element in the first exon of the maize Shrunken-1 gene with the following intron 1 enhances reporter gene expression up to 1000-fold. Plant Mol Biol 16: 199–207 (1991).
Mozer TJ: Control of protein synthesis in barley aleurone layers by the plant hormones gibberellic acid and abscisic acid. Cell 20: 479–485 (1980).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15: 473–479 (1962).
Murphy G, Kavanagh TA: Speeding-up the sequencing of double stranded DNA. Nucl Acids Res 16: 5198 (1988).
Rogers JC, Dean D, Heck GR: Aleurain: A barley thiol protease closely related to mammalian cathepsin H. Proc Natl Acad Sci USA 82: 6512–6516 (1985).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1989).
Skriver K, Olsen FL, Rogers JC, Mundy J: Cis-acting DNA elements responsive to gibberellin and its antagonist abscisic acid. Proc Natl Acad Sci USA 88: 7266–7270 (1991).
Watanabe H, Abe K, Emori Y, Hosoyama H, Arai S: Molecular cloning and gibberellin-induced expression of multiple cysteine proteases of rice seeds (oryzains). J Biol Chem 266: 16897–16902 (1991).
Whittier RF, Dean AD, Rogers JC: Nucleotide sequence analysis of alpha-amylase and thiol protease genes that are hormonally regulated in barley aleurone cells. Nucl Acids Res 15: 2515–2535 (1987).
Zwar JA, Hooley R: Hormonal regulation of α-amylase gene transcription in wild oat (Avena fatua L.) aleurone protoplasts. Plant Physiol 80: 459–43 (1986).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Cejudo, F.J., Ghose, T.K., Stabel, P. et al. Analysis of the gibberellin-responsive promoter of a cathepsin B-like gene from wheat. Plant Mol Biol 20, 849–856 (1992). https://doi.org/10.1007/BF00027156
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00027156