Abstract
Upstream regulatory sequences (URS) of the gene that encodes the α′ subunit of β-conglycinin, the 7S soybean seed storage protein, includes two RY repeat elements. The role of RY elements and sequences that bind soybean embryo factors 3 and 4 (SEF3 and SEF4; Allen et al., Plant Cell 1 (1989) 623–631) in regulating expression of the promoter was studied following site directed mutagenesis. Specific mutations introduced into these sequences abolished the in vitro binding activities of SEF3 and SEF4. The biological activities resulting from the mutations were determined in transgenic plants using two chimeric promoters comprising sequences from the CaMV 35S promoter and the α′ subunit promoter. The uidA reporter gene was used to assess the levels of gene expression in transgenic plants. The mutations in the RY element and SEF3 and SEF4 binding sites had little effect on expression of the α′ promoter. By contrast, mutations in the RY element had significant effect on gene expression when the URS from the α′ promoter was ligated upstream of the core 35S promoter. Mutations in the RY element abolished the seed specific enhancing activity of the α′ URS and caused expression of the chimeric promoter in leaves. These results indicate that the RY element plays a key role in seed-specific gene regulation in coordination with other cis-acting elements.
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References
Allen RD, Bernier F, Lessard PA, Beachy RN: Nuclear factors interact with a soybean β-conglycinin promoter. Plant Cell 1: 623–631 (1989).
Beachy RN, Chen Z-L, Horsch RB, Rogers SG, Hoffmann NJ, Fraley RT: Accumulation and assembly of soybean β-conglycinin in seeds of transformed petunia plants. EMBO J 4: 3047–3053 (1985).
Bäumlein H, Boerjan W, Nagy I, Panitz R, Inzé D, Wobus U: Upstream sequences regulating legumin gene expression in heterologous transgenic plants. Mol Gen Genet 225: 121–128 (1991).
Bäumlein H, Nagy I, Villarroel R, Inzé D, Wobus U: 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 (1992).
Benfey PN, Ren L, Chua N-H: The CaMV 35S enhancer contains at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J 8: 2195–2201 (1989).
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).
Burrow MD, Sen P, Chlan CA, Murai N: 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 (1992).
Bustos MM, Begum D, Kalkan FA, Battraw MJ, Hall TC: 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 10: 1469–1479 (1991).
Chamberland S, Daigle N, Bernier F: The legumin boxes and the part of a soybean β-conglycinin promoter are involved in seed gene expression in transgenic tobacco plants. Plant Mol Biol 19: 937–949 (1992).
Chen Z-L, Pan NS, Beachy RN: A DNA sequence element that confers seed-specific enhancement of a constitutive promoter. EMBO J 7: 297–302 (1988).
Chen Z-L, Schuler MA, Beachy RN: Functional analysis of regulatory elements in a plant embryo specific gene. Proc Natl Acad Sci USA 83: 8560–8564 (1986).
Dickinson CD, Evans PR, Nielsen NC: RY repeats are conserved in the 5′ flanking regions of legume seed protein genes. Nucl Acids Res 16: 371 (1988).
Figurski DH, Helinski DR: Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76: 1648–1652 (1979).
Fraley RT, Rogers SG, Horsch RB, Eichholtz DA, Flick JS, Fink CL, Hoffman NL, Sanders PR: The SEV system: a new disarmed Ti plasmid vector system for plant transformation. Bio/technology 3: 629–635 (1985).
Fujiwara T: Seed specific expression of soybean storage protein genes. Ph.D. thesis, University of Tokyo (1992).
Fujiwara T, Beachy RN: Expression of soybean seed storage protein genes in transgenic plants; their effects on expression of a neighboring gene and position dependency. Plant Cell Physiol 34: 13–20 (1993).
Fujiwara T, Lessard PA, Beachy RN: Seed specific repression of GUS activity in tobacco plants by antisense RNA. Plant Mol Biol 20: 1059–1069 (1992).
Gayler KR, Sykes GE: β-Conglycinins in developing soybean seeds. Plant Physiol 67: 958–961 (1981).
Horsch RB, Klee HJ: Rapid assay of foreign gene expression in leaf discs transformed by Agrobacterium tumefaciens: role of T-DNA borders in the transfer process. Proc Natl Acad Sci USA 83: 4428–4432 (1986).
Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).
Katagiri F, Lam E, Chua N-H: Two tobacco DNA-binding proteins with homology to the nuclear factor CREB. Nature 340: 727–730 (1989).
Kawagoe Y, Murai N: 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 (1992).
Kunkel TA, Roberts JD, Zakour RA: Rapid and efficient site-specific mutagenesis without phenotypic selection. Meth Enzymol 154: 367–382 (1987).
Lam E, Kano-Murakami Y, Gilmartin P, Niner B, Chua N-H: A metal-dependent DNA-binding protein interacts with a constitutive element of a light-responsive promoter. Plant Cell 2: 857–866 (1990).
Lelievre J-M, Oliveira LO, Nielsen NC: 5′-CATGCAT-3′ elements modulate the expression of glycinin genes. Plant Physiol 98: 387–391 (1992).
Lessard PA, Allen RD, Bernier F, Crispino JD, Fujiwara T, Beachy RN: Multiple nuclear factors interact with upstream sequence of differentially regulated β-conglycinin genes. Plant Mol Biol 16: 397–413 (1991).
Lessard PA, Fujiwara T, Allen RD, Beachy RN: Analysis of cis acting elements from genes encoding β-conglycinins. J Cell Biochem Suppl 15A: 100 (Abstract A441) (1991).
Lewin B: Genes, 4th ed., pp. 84–85. Oxford University Press, New York (1990).
Matzke AJM, Stöger EM, Schernthaner JP, Matzke MA: Deletion analysis of a zein gene promoter in transgenic tobacco plants. Plant Mol Biol 14: 323–332 (1990).
Meinke DW, Chen J, Beachy RN: Expression of storage-protein genes during soybean seed development. Planta 153: 130–139 (1981).
Naito S, Dubé PH, Beachy RN: Differential expression of β-conglycinin α′ and β subunit genes in transgenic plants. Plant Mol Biol 11: 109–123 (1988).
Rogers ST, Horsch RB, Fraley RT: Gene transfer in plants: production of transformed plants using Ti plasmid vectors. Meth Enzymol 118: 627–640 (1986).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1989).
Shortwell MA, Larkins BA. The biochemistry and molecular biology of seed storage proteins. In: The biochemistry of plants, vol 15, pp. 297–345. Academic Press, New York (1989).
Vieira J, Messing J: Production of single-stranded plasmid DNA. Meth Enzymol 153: 3–11 (1987).
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Fujiwara, T., Beachy, R.N. Tissue-specific and temporal regulation of a β-conglycinin gene: roles of the RY repeat and other cis-acting elements. Plant Mol Biol 24, 261–272 (1994). https://doi.org/10.1007/BF00020166
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DOI: https://doi.org/10.1007/BF00020166