Abstract
Nuclear proteins from bean (Phaseolus vulgarus) embryos bind specifically to a 55 bp DNA sequence located upstream of the seed storage protein gene phaseolin. This sequence is capable of elevating gene expression in transgenic tobacco plants by as much as 150-fold when fused to a chimeric β-glucuronidase reporter gene. Results presented in this paper demonstrate that nuclear extracts from carrot embryos bind to a phaseolin DNA sequence that includes a phaseolin activator sequence. This specific DNA binding activity is modulated during somatic embryogenesis. Two separable protein species react specifically with the labeled phaseolin DNA fragment (58.0 and 51.7 dDa). These results suggest that the cis- and trans-acting elements controlling gene expression have been highly conserved during evolution.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- bp:
-
base pairs
- CAMV:
-
cauliflower mosaic virus
- GUS:
-
β-glucuronidase
- kDa:
-
kilodalton
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
References
Bustos MM, Guiltinan MJ, Jordano J, Begum D, Kalkan FA, Hall TC: Regulation of GUS expression in transgenic tobacco plants by an A/T-rich cis-acting sequence found upstream of a French bean β-phaseolin gene. The Plant Cell (1989) in press.
Choi JC, Liu L, Borkird C, Sung ZR: Cloning of genes developmentally regulated during plant embryogenesis. Proc Natl Acad Sci USA: 84: 1906–1910 (1987).
Cyr RJ, Bustos MM, Guiltinan MJ, Fosket DE: Developmental modulation of tubulin protein and mRNA levels during somatic embryogenesis in cultured carrot cells. Planta 171: 365–376 (1987).
DeVries S, Booij H, Meyerink P, Huisman G, vanSpanje M, Wilde HD, Thomas TL, vanKammen A: Acquisition of embryogenic potential in carrot cell suspension cultures. Planta 176: 196–204 (1988).
Ferl RJ, Nick HS: In vivo detection of regulatory factor binding sites in the 5′ flanking region of maize Adh. 1, J Biol Chem 262: 7947–7950 (1987).
Guiltinan MJ, Klassy RC, Hall TC. Functional analysis of a protein binding domain from the 5′ region of the phaseolin gene. The second international congress of Plant Molecular Biology: abs. 46 (1988).
Guiltinan MJ, Schelling ME, Ehtesham NZ, Thomas JC, Christensen ME: The nucleolar RNA-binding protein B-36 is highly conserved among plants. Eur J Cell Biol 46: 547–553 (1988).
Green PJ, Kay SA, Chua N-H: Sequence-specific interactions of a pea nuclear factor with light-responsive elements upstream of the rbc-S3A gene. EMBO J 6: 2543–2549 (1987).
Halperin W: Alternative morphogenetic events in cell suspensions. Am J Bot 53: 443–453 (1966).
Jensen EO, Marcker KA, Schell J, Bruijn FJ: Interaction of a nodule specific trans-acting factor with distinct DNA elements in the soybean leghaemoglobin lbc3 upstream region. EMBO J 7: 1265–1271 (1988).
Jofuku KD, Okamura JK, Goldberg RB: Interaction of an embryo DNA binding protein with a soybean lectin gene upstream region. Nature 328: 734–737 (1997).
Keller GL, Nikolau BJ, Ulrich TH, Wurtele ES: Comparison of starch and ADP-glucose pyrophosphorylase levels in nonembryogenic cells and developing embryos from induced carrot cultures. Plant Physiol 86: 451–456 (1988).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).
Maier UG, Brown JWS, Toloczyki C, Feix G: Binding of a nuclear factor to a consensus sequence in the 5′ flanking region of zein genes from maize. EMBO J 6: 17–22 (1987).
Maier UG, Brown JWS, Schmitz LM, Schwall M, Dietrich G, Feix G: Mapping of tissue-dependent and independent protein binding sites to the 5′ region of a zein gene. Mol Gen Genet 212: 241–245 (1988).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Maniatis T, Goodbourn S, Fischer FA: Regulation of inducible and tissue-specific gene expression. Science 236: 1237–1245 (1987).
Miskimins WK, Roberts MP, McClellan A, Ruddle FH: Use of a protein-blotting procedure and a specific DNA probe to identify nuclear proteins that recognize the promoter region of the transferrin receptor gene. Proc Natl Acad Sci USA 82: 6741–6744 (1985).
Oakley BR, Kirsh DR, Morris NR: A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem 105: 361–363 (1980).
Sengupta-Gopalan C, Reichert NA, Barker RF, Hall TC: Developmentally regulated expression of the bean β-phaseolin gene in tobacco seed. Proc Natl Acad Sci USA 82: 3320–3324 (1985).
Slightom JL, Sun SM, Hall TC: Complete nucleotide sequence of a French bean storage protein gene: Phaseolin. Proc Natl Acad Sci USA 80: 1897–1901 (1983).
Smith JA, Choi JH, Krass MA, Karu AE, Sung ZR: Monoclonal antibodies that recognize proteins unique to somatic embryos of Daucus carota. In Terzi M, Pitto L Sung ZR (eds) Proceedings of Somatic Embryogenesis Workshop (San Miniato, Italy), pp. 86–94 (1985).
Steward FC, Mapes MO, Kent AE, Holsten RD: Growth and development of cultured plant cells. Science 143: 20–27 (1964).
Sung ZR, Okimoto R: Embryonic proteins in somatic embryos of carrots. Proc Natl Acad Sci USA 78: 3683–3687 (1981).
Sung ZR, Okimoto R: Coordinate gene expression during somatic embryogenesis in carrot. Proc Natl Acad Sci USA 80: 2661–2665 (1983).
Thomas JC, Guiltinan MJ, Thomas TT, Nessler C: Agrobacterium tumefaciens transformation of Dacus carota hypocotyl sections. Plant Molecular Biology (1989) in press.
Werr W, Springer B, Schurmann J, Bellman R: Multiple interactions between nuclear proteins of Zea mays and the promoter of the Shrunken gene. Mol Gen Genet 212: 342–350 (1988).
Wilde HD, Nelson WS, Booij H, DeVries S, Thomas TL: Gene expression programs in embryogenic carrot cultures. Planta 176: 205–211 (1988).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Guiltinan, M.J., Thomas, J.C., Nessler, C.L. et al. Expression of DNA binding proteins in carrot somatic embryos that specifically interact with a cis regulatory element of the French bean phaseolin gene. Plant Mol Biol 13, 605–610 (1989). https://doi.org/10.1007/BF00027322
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00027322