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Differences in cell type-specific expression of the gene Opaque 2 in maize and transgenic tobacco

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Abstract

The Opaque 2 (O2) gene encodes a transcriptional activator of the basic region/leucine zipper family, which controls the synthesis of a major storage protein class in maize endosperm, the 22 kDa α-zeins, and of several other non-zein polypeptides including b32. We demonstrate, by analysing O2 mRNAs in different organs of maize plants, that the O2 gene is only active in the endosperm. Its transcription is precisely controlled during seed development: O2 mRNAs are first detected 10 days after pollination and accumulate in the endosperm over a period of 20 days. When introduced into tobacco plants, the O2 promoter directs the expression of the β-glucuronidase (GUS) reporter gene in endosperm, but also in the embryo, cotyledons and pollen. The first 185 by of the O2 promoter is sufficient for developmentally regulated expression in tobacco seeds. A distinct cis-acting element, located between positions −185 and −520, directs expression in the cotyledons of tobacco seedlings. The possible origins of this breakdown in promoter specificity in the heterologous host are discussed.

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References

  • Bartels D, Thompson RD (1983) The characterization of cDNA clones coding for wheat storage proteins. Nucleic Acids Res 10:2961–2975

    Google Scholar 

  • Bass HW, Webster C, O'Brian G, Roberts JKM, Boston RS (1992) A maize ribosome-inactivating protein is controlled by the transcriptional activator opaque-2. Plant Cell 4:225–234

    Google Scholar 

  • Bäumlein H, Wobus U, Pustell J, Kafatos FC (1986) The legumin gene family: structure of a B-type gene of Vicia faba and a possible legumin gene-specific regulatory element. Nucleic Acids Res 14:2707–2720

    Google Scholar 

  • Bäumlein H, Boerjan W, Nagy I, Panitz R, Inze D, Wobus U (1991) Upstream sequences regulating legumin gene expression in heterologous transgenic plants. Mol Gen Genet 225:121–128

    Google Scholar 

  • Bäumlein H, Nagy I, Villarroel R, Inze D, Wobus U (1992) Cis-analysis of a seed protein gene promoter: the conservative RY repeat CATGCATG within the legumin box is essential for tissue-specific expression of a legumin gene. Plant J 2:233–239

    Google Scholar 

  • Benfey PN, Ren L, Chua NH (1989) The CaMV 35S enhancer contains at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J 8:2195–2202

    Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Burow MD, Sen P, Shlan CA, Murai N (1992) Developmental control of the β-phaseolin gene requires positive, negative, and temporal seed-specific transcriptional regulatory elements and a negative element for stem and root expression. Plant J 2:537–548

    Google Scholar 

  • Bustos MM, Begum D, Kalkan FA, Battraw MJ, Hall TC (1991) Positive and negative cis-acting DNA domains are required for spatial and temporal regulation of gene expression by a seed storage protein promoter. EMBO J 6:1469–1479

    Google Scholar 

  • Bustos MM, Guiltinan MJ, Jordano J, Begum D, Kalkan FA, Hall TC (1989) Regulation of β-glucuronidase expression in transgenic tobacco plants by an A/T-rich, cis-acting sequence found upstream of a french bean β-phaseolin gene. Plant Cell 1:839–853

    Google Scholar 

  • Colot V, Robert LS, Kavanagh TA, Bevan MW, Thompson RD (1987) Localization of sequences in wheat endosperm protein genes which confer tissue-specific expression in tobacco. EMBO J 6:3559–3564

    Google Scholar 

  • Dickinson CD, Evans RP, Nielsen NC (1988) RY repeats are conserved in the 5′ flanking regions of legume seed-protein genes. Nucleic Acids Res 16:371

    Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Giovinazzo G, Manzocchi LA, Bianchi MW, Coraggio I, Viotti A (1992) Functional analysis of the regulatory region of a zein gene in transiently transformed protoplasts: Plant Mol Biol 19:257–263

    Google Scholar 

  • Goldberg RB, Barker SJ, Perez-Grau L (1989) Regulation of gene expression during plant embryogenesis. Cell 56:149–160

    Google Scholar 

  • Hagen G, Rubinstein I (1981) Complex organization of zein proteins in maize. Gene 13:239–249

    Google Scholar 

  • Hammond-Kosack 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 

  • Hartings H, Maddaloni M, Lazzaroni N, Di Fonzo N, Motto M, Salamini F, Thompson R (1989) The O2 gene, which regulates zein deposition in maize endosperm encodes a protein with structural homologies to transcriptional activators. EMBO J 8:2795–2801

    Google Scholar 

  • Hattori T, Vasil V, Rosenkrans L, Curtis Hannah L, McCarty DR, Vasil IK (1992) The Vivaparous-1 gene and abscisic acid activate the C1 regulatory gene for anthocyanin biosynthesis during seed maturation in maize. Genes Devel 6:609–618

    Google Scholar 

  • Hoekema A, Hirsch PR, Hooykass PJJ, Schilperoort RA (1983) A binary plant vector strategy based on the separation of virand T-region of the Agrobacterium tumefaciens Ti plasmid. Nature 303:179–180

    Google Scholar 

  • Höfgen R, Willmitzer L (1988) Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res 16:9877

    Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1230

    Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBOJ 6:3901–3907

    Google Scholar 

  • Jones RA, Larkins BA, Tsai CY (1977a) Storage protein synthesis in maize. III. Developmental changes in membrane-bound polyribosome composition and in vitro protein synthesis of normal and opaque-2 maize. Plant Physiol 59:733–737

    Google Scholar 

  • Jones RA, Larkins BA, Tsai CY (1977b) Storage protein synthesis in maize. II. Reduced synthesis of a major zein component by the opaque-2 mutant of maize. Plant Physiol 59:525–529

    Google Scholar 

  • Katagiri F, Chua NH (1992) Plant transcriptional activators: present knowledge and future challenges. Trends Genet 8:22–27

    Google Scholar 

  • Kawagoe Y, Murai N (1992) Four distinct nuclear proteins recognize in vitro the proximal promoter of the bean seed storage protein β-phaseolin gene conferring spatial and temporal control. Plant J 2:927–936

    Google Scholar 

  • Kawalleck P (1991) Resistance to pathogens in plants: identification of genes whose products participate in induced defence reactions in parsley. PhD thesis, University of Cologne

  • Kodrzycki R, Boston RS, Larkins BA (1989) The opaque-2 mutation of maize differentially reduces zein gene transcription. Plant Cell 1:104–114

    Google Scholar 

  • Kowles RV, Phillips R (1990) Endosperm development in maize. Int Rev Cytol 112:97–135

    Google Scholar 

  • Lam E, Chua NH (1991) Tetramer of a 21-base pair synthetic element confers seed expression and transcriptional enhancement in response to water stress and abscisic acid. J Biol Chem 26:17131–17135

    Google Scholar 

  • Lelièvre JM, Oliveira LO, Nielsen NC (1992) 5′-CATGCAT-3′ elements modulate the expression of glycinin genes. Plant Physiol 98:387–391

    Google Scholar 

  • Lohmer S, Maddaloni M, Motto M, Di Fonzo N, Hartings H, Salamini F, Thompson RD (1991) The maize regulatory locus Opaque-2 encodes a DNA-binding protein which activates the transcription of the b-32 gene. EMBO J 10:617–624

    Google Scholar 

  • Lohmer S, Maddaloni M, Motto M, Salamini F, Thompson RD (1993) Translation of the mRNA of the maize transcriptional activator Opaque-2 is inhibited by upstream open reading frames present in the leader sequence. Plant Cell 5:65–73

    Google Scholar 

  • Luan S, Bogorad L (1992) A rice cab gene promoter contains separate cis-acting elements that regulate expression in dicot and monocot plants. Plant Cell 4:971–981

    Google Scholar 

  • Maier UG, Brown JWS, Toloczyki C, Feix G (1987) Binding of a nuclear factor to a consensus sequence in the 5′ flanking region of zein genes from maize. EMBO J 6:17–22

    Google Scholar 

  • Manzocchi LA, Bianchi MW, Viotti A (1989) Expression of zein in long term cultures of wild type and opaque-2 maize endosperm. Plant Cell Rep 7:639–643

    Google Scholar 

  • Marris C, Gallois P, Copley J, Kreis M (1988) The 5′ flanking region of a barley B hordein gene controls tissue and developmental specific CAT expression tobacco plants. Plant Mol Biol 10:359–366

    Google Scholar 

  • Matzke AJM, Stöger EM, Schernthaner JP, Matzke MA (1990) Deletion analysis of a zein gene promoter in transgenic tobacco plants. Plant Mol Biol 14:323–332

    Google Scholar 

  • Motto M, Maddaloni M, Ponziani G, Brembilla M, Marotta R, Di Fonzo N, Soave C, Thompson RD, Salamini F (1988) Molecular cloning of the o2m5 allele of Zea mays using transposon marking. Mol Gen Genet 212:488–494

    Google Scholar 

  • Motto M, Di Fonzo N, Hartings H, Maddaloni M, Salamini F, Soave C, Thompson RD (1989) Regulatory genes affecting maize storage protein synthesis. Oxf Surv Plant Mol Cell Biol 6:87–114

    Google Scholar 

  • Murphy J, Dalby A (1971) Changes in the protein fractions of developing normal and opaque-2 maize endosperm. Cereal Chem 48:336–349

    Google Scholar 

  • Paz-Ares J, Ghosal D, Wienand U, Peteson PA, Saedler H (1987) The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators. EMBO J 6:3553–3558

    Google Scholar 

  • Pysh DL, Aukerman MJ, Schmidt RJ (1993) OHP1: a maize basic domain/leucine zipper protein that interacts with opaque2. Plant Cell 5:227–236

    Google Scholar 

  • Quayle T, Feix G (1992) Functional analysis of the −300 region of maize zein genes. Mol Gen Genet 231:369–374

    Google Scholar 

  • Roberts LS, Thompson RD, Flavell RB (1989) Tissue-specific expression of a wheat high molecular weight glutenin gene in transgenic tobacco. Plant Cell 1:569–578

    Google Scholar 

  • Salinas J, Oeda K, Chua NH (1992) Two G-box-related sequences confer different expression patterns in transgenic tobacco. Plant Cell 4:1485–1493

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Schäffner AR, Sheen J (1991) Maize rbcs promoter activity depends on sequence elements not found in dicots rbcs promoters. Plant Cell 3:997–1012

    Google Scholar 

  • Schernthaner JP, Matzke MA, Matzke AJM (1988) Endospermspecific activity of a zein gene promoter in transgenic tobacco plants. EMBO J 5:1249–1255

    Google Scholar 

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

    Google Scholar 

  • Schmidt RJ, Burr FA, Aukerman MJ, Burr B (1990) Maize regulatory gene Opaque-2 encodes a protein with a “leucine-zipper” motif that binds to zein DNA. Proc Natl Acad Sci USA 87:46–50

    Google Scholar 

  • Schmidt RJ, Ketudat M, Aukerman MJ, Hoschek G (1992) Opaque-2 is a transcriptional activator that recognizes a specific target site in 22-kD zein genes. Plant Cell 4:689–700

    Google Scholar 

  • Sengupta-Gopalan C, Reichert NA, Barker RF, Hall TC, Kemp JD (1985) Developmentally regulated expression of the bean β-phaseolin gene in tobacco seed. Proc Natl Acad Sci USA 82:3320–3324

    Google Scholar 

  • Soave C, Tardani L (1981) Zein level in maize endosperm depends on a protein under control of the Opaque-2 and Opaque-6 loci. Cell 27:403–410

    Google Scholar 

  • Takaiwa F, Oono K, Kato A (1991) Analysis of the 5′ flanking region responsible for the endosperm-specific expression of a rice glutelin chimeric gene in transgenic tobacco. Plant Mol Biol 16:49–58

    Google Scholar 

  • Thomas MS, Flavell RB (1990) Identification of an enhancer element for the endosperm-specific expression of high molecular weight glutenin. Plant Cell 2:1171–1180

    Google Scholar 

  • Ueda T, Messing J (1991) A homologous expression system for cloned zein genes. Theor Appl Genet 82:93–100

    Google Scholar 

  • Ueda T, Waverczak W, Ward K, Sher N, Ketudat M, Schmidt RJ, Messing J (1992) Mutations of the 22- and 27-kD zein promoters affect transactivation by the opaque-2 protein. Plant Cell 4:701–709

    Google Scholar 

  • Uknes S, Dincher S, Friedrich L, Negrotto D, Williams S, Thompson-Taylor H, Potter S, Ward E, Ryals J (1993) Regulation of pathogenesis-related protein-1a gene expression in tobacco. Plant Cell 5:159–169

    Google Scholar 

  • Wilson DR, Larkins BA (1984) Zein gene organization in maize and related grasses. J Mol Evol 20:330–340

    Google Scholar 

  • Zheng Z, Kawagoe Y, Xiao S, Okita T, Hau TL, Lin A, Murai N (1993) 5′ distal and proximal cis-acting regulator elements are required for developmental control of a rice seed storage protein glutelin gene. Plant Cell 4:357–366

    Google Scholar 

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Authors and Affiliations

  1. Max-Planck-Institut für Züchtungsforschung, Carl-von-Linné-Weg 10, D-50829, Köln, Germany

    P. Gallusci, F. Salamini & R. D. Thompson

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  1. P. Gallusci
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  2. F. Salamini
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  3. R. D. Thompson
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Communicated by J. Schell

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Gallusci, P., Salamini, F. & Thompson, R.D. Differences in cell type-specific expression of the gene Opaque 2 in maize and transgenic tobacco. Molec. Gen. Genet. 244, 391–400 (1994). https://doi.org/10.1007/BF00286691

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  • DOI: https://doi.org/10.1007/BF00286691

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