Skip to main content
Log in

Disruption of an overlapping E-box/ABRE motif abolished high transcription of the napA storage-protein promoter in transgenic Brassica napus seeds

  • Published:
Planta Aims and scope Submit manuscript

Abstract

The storage protein napin is one of the major protein components of Brassica napus L. (oilseed rape) seeds. To investigate the transcriptional regulation of the napin promoter, different constructs of the napin gene napA promoter were fused to the Escherichia coli uidA gene and transformed into B. napus. A -152-bp promoter construct directed a strong expression of the marker gene in mature seeds. The 5′ deletion of an additional 8 completely abolished this activity. This deletion disrupted sequence motifs that are similar to an E-box, (CA↓ NNTG) and an ABRE (CGCCA↓CGTGTCC) element (identity is indicated by bold face). Further, internal deletion of a segment corresponding to -133 to -121 caused an eightfold reduction in the activity of the -152 construct. This region contains an element, CAAACAC, conserved in many storage-protein gene promoters. These results imply that the E-box/ABRE-like sequence is a major motif of the napA promoter and suggest that the CAAACAC sequence is important for high activity of the napA promoter. Similar results have been obtained by analysing some of the constructs in transgenic tobacco, suggesting that many of the cis-elements in the napA promoter are conserved, at least in dicotyledonous species.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

ABA:

abscisic acid

GUS:

glucuronidase

References

  • Bäumlein H, Miséra S, Luersßen H, Kölle K, Horstman C, Wobus U, Müller AJ (1994) The FUS3 gene of Arabidopsis thaliana is a regulator of gene expression during late embryogenesis. Plant J6: 379–387

    Google Scholar 

  • Blundy KS, Blundy MAC, Crouch ML (1991) Differential expression of members of the napin storage protein gene family during embryogenesis in Brassica napus. Plant Mol Biol 17: 1099–1104

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye-binding. Anal Biochem 72: 248–254

    Article  CAS  PubMed  Google Scholar 

  • De Block M, De Brouwer D, Tenning P (1989) Transformation of Brassica napus and Brassica oleracea using Agrobacterium tumefaciens and the expression of the bar and neo genes in transgenic plants. Plant Physiol 91: 694–701

    Google Scholar 

  • DeLisle AJ, Crouch ML (1989) Seed storage protein transcription and mRNA levels in Brassica napus during development and in response to exogenous abscisic acid. Plant Physiol 91: 617–623

    Google Scholar 

  • Finkelstein RR, Somerville CR (1990) Three classes of abscisic acid (ABA)-insensitive mutations of Arabidopsis define genes that control overlapping subset of ABA responses. Plant Physiol 94: 1172–1179

    Google Scholar 

  • Gallusci P, Salamini F, Thompson RD (1994) Differences in cell type-specific expression of the Opaque2 in maize and transgenic tobacco. Mol Gen Genet 244: 391–400

    Google Scholar 

  • Giraudat J, Parcy F, Bertauche N, Gost F, Leung J, Morris P-C, Bouvier-Durand M, Vartanian N (1994) Current advances in abscisic acid action and signalling. Plant Mol Biol 26: 1557–1577

    Google Scholar 

  • Grossi de Sa MF, Weinberg DF, Rech EL, Barros LMG, Aragoa FJL, Holmstroem K-O, Gander ES (1994) Functional studies on a seed-specific promoter from a Brazil nut gene. Plant Sci 103: 189–198

    Google Scholar 

  • Guerche P, Tire C, Grossi de Sa F, De Clercq A, Van Montagu M, Krebbers E (1990) Differential expression of the Arabidopsis 2S albumin genes and the effect of increasing gene family size. Plant Cell 2: 469–478

    Google Scholar 

  • Guiltinan MJ, Marcotte WR Jr, Quatrano RS (1990) A plant leucin zipper protein recognizes an abscisic acid response element. Science 250: 267–271

    Google Scholar 

  • Gustavsson H-O, Ellerström M, Stålberg K, Ezcurra I, Koman A, Höglund A-S, Rask L, Josefsson L-G (1991) Distinct sequence elements in a napin promoter interacts in vitro with DNA-binding proteins from Brassica napus. Physiol Plant 82: 205–212

    Google Scholar 

  • Hammond-Kozack MCU, Holdsworth MJ, Bevan MW (1993) In vivo footprinting of a low molecular weight glutenin gene (LMWG-1D1) in wheat endosperm. EMBO J 12: 545–554

    Google Scholar 

  • Höglund A-S, Rödin J, Larsson E, Rask L (1991) The distribution of napin and cruciferin in developing rape seed embryos. Plant Physiol 98: 509–515

    Google Scholar 

  • Izawa T, Foster R, Chua N-H (1993) Plant bZIP protein DNA binding specificity. J Mol Biol 230: 1131–1144

    Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep 5: 387–405

    CAS  Google Scholar 

  • Josefsson L-G, Lenman M, Ericson ML, Rask L (1987) Structure of a gene encoding the 1.7S storage protein, napin, from Brassica napus. J Biol Chem 262: 12196–12201

    Google Scholar 

  • Kawagoe Y, Campell BR, Murai N (1994) Synergism between CACGTG (G-box) and CACCTG cis-elements is required for activation of the bean seed storage protein β-phaseolin gene. Plant J 5: 885–890

    Google Scholar 

  • Kodrzycki R, Boston RS, Larkins BA (1989) The opaque-2 mutation of maize differentially reduces zein gene expression. Plant Cell 1: 105–114

    Google Scholar 

  • Koorneef M, Reuling G, Karssen CM (1984) The isolation and characterization of abscisic acid-insensitive mutants of Arabidopsis thaliana. Physiol Plant 61: 385–393

    Google Scholar 

  • Krebbers E, Herdies L, De Clercq A, Seurinick J, Leemans J, van Damme J, Segura M, Geysen G, van Montagu M, Vandekerckhove J (1988) Determination of the processing sites of an Arabidopsis 2S albumin and characterization of the complete gene family. Plant Physiol 87: 859–866

    Google Scholar 

  • Kriz A, Wallace MS, Paiva R (1990) Globulin gene expression in embryos of maize viviparous mutants. Plant Physiol 92: 538–542

    Google Scholar 

  • Mettler U (1987) A simple and rapid method for minipreparation of DNA from tissue cultured plant cells. Plant Mol Biol Rep 5: 346–349

    Google Scholar 

  • McCarty DR, Hattori T, Carson CB, Vasil V, Lazar M, Vasil IK (1991) The viviparous-1 developmental gene of maize encodes a novel transcriptional activator. Cell 66: 895–905

    Google Scholar 

  • Morton RL, Quiggin D, Higgins TJV (1995) Regulation of seed storage protein gene expression. In: Kigel J, Galili G (eds) Seed development and germination. Marcel Dekker, Inc. New York, Basel, Hongkong, pp 103–136

    Google Scholar 

  • Parcy F, Valon C, Raynal M, Gaubier-Comella P, Delseny M, Giraudat J (1994) Regulation of gene expression programs during arabidopsis seed development: Roles of the AB13 locus and endogenous abscisic acid. Plant Cell 6: 1567–1582

    Google Scholar 

  • de Pater S, Katagiri F, Kijne J, Chua N-H (1994) bZIP proteins bind to a palindromic sequence without an ACGT core located in a seed-specifying element of the pea lectin promoter. Plant J 6: 133–140

    Google Scholar 

  • Pysh LD, Aukerman MJ, Schmidt RJ (1993) OHP1: A maize basic domain/leucin zipper protein that interacts with opaque 2. Plant Cell 5: 227–236

    Google Scholar 

  • Sablowski RWM, Moyano E, Culianez-Macia FA, Schuch W, Martin C, Bevan M (1994) A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes. EMBO J 13: 128–137

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989): Molecular cloning, A laboratory manual, second edition. Cold Spring Harbor Laboratory Press, New York, USA

    Google Scholar 

  • Schindler U, Menkens AE, Beckmann H, Ecker JR, Cashmore AR (1992) Heterodimerization between light-regulated and ubiquitously expressed Arabidopsis GBF bZIP proteins. EMBO J 11: 1261–1274

    Google Scholar 

  • Schmidt RJ, Burr FA, Burr B (1987) Transposon tagging and molecular analysis of the maize regulatory locus Opaque-2. Science 238: 960–963

    Google Scholar 

  • Stålberg K, Ellerström M, Josefsson L-G, Rask L (1993) Deletion analysis of a 2S seed storage protein promoter of Brassica napus in transgenic tobacco. Plant Mol Biol 23: 671–683

    Google Scholar 

  • Thomas TL (1993) Gene expression during plant embryogenesis and germination: An overview. Plant Cell 5: 1401–1410

    Google Scholar 

  • Yunes JA, Cord Neto G, Da Silva MJ, Leite A, Ottoboni LMM, Arruda P (1994) The transcriptional activator opaque2 recognize two different target sequences in the 22-kD-like a-prolamin genes. Plant Cell 6: 236–249

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kjell Stålberg.

Additional information

Dr. Enno Krebbers, (Plant Genetic Systems N.V., Gent, Belgium) kindly provided the Agrobacterium strain C58C1(PMP90). Bertil Blom and Elisabeth Westergren are gratefully acknowledged for expert technical assistance. This work was supported by grants form the Swedish Natural Science Research Council, The Swedish Research Council for Forestry and Agriculture, The Swedish Institute and the Nilsson-Ehle Foundation.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stålberg, K., Ellerstöm, M., Ezcurra, I. et al. Disruption of an overlapping E-box/ABRE motif abolished high transcription of the napA storage-protein promoter in transgenic Brassica napus seeds. Planta 199, 515–519 (1996). https://doi.org/10.1007/BF00195181

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00195181

Key words

Navigation