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. 1989 Sep;1(9):855–866. doi: 10.1105/tpc.1.9.855

A sunflower helianthinin gene upstream sequence ensemble contains an enhancer and sites of nuclear protein interaction.

J Jordano 1, C Almoguera 1, T L Thomas 1
PMCID: PMC159822  PMID: 2535527

Abstract

Genes encoding helianthinin, the major seed protein in sunflower, are highly regulated. We have identified putative cis-acting and trans-acting elements that may function in the control of helianthinin expression. A 404-base pair DNA fragment on the sunflower helianthinin gene HaG3D, located 322 base pairs from the transcriptional start site, enhanced beta-glucuronidase expression in transgenic tobacco embryos. Sequences within this fragment were found to bind nuclear proteins present in both sunflower embryo and hypocotyl nuclear extracts. The binding site was localized by phenanthroline-copper ion footprinting experiments to A/T-rich sequences located from -705 to -654. Binding competition experiments revealed that these sunflower proteins also bind to upstream promoter sequences from another helianthinin gene (HaG3A) and two other plant embryo-specific genes, carrot DcG3 and French bean phaseolin. However, sequences of the cauliflower mosaic virus 35S promoter/enhancer complex failed to compete for its binding. Phenanthroline-copper ion footprinting experiments showed that the binding sites for the sunflower proteins in HaG3A (-1463 to -1514 and -702 to -653) and in phaseolin (-671 to -627) are also very A/T-rich, have similar sizes, and are located at similar distances from their respective promoters.

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Selected References

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  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Bustos M. M., Guiltinan M. J., Jordano J., Begum D., Kalkan F. A., Hall T. C. Regulation of beta-glucuronidase expression in transgenic tobacco plants by an A/T-rich, cis-acting sequence found upstream of a French bean beta-phaseolin gene. Plant Cell. 1989 Sep;1(9):839–853. doi: 10.1105/tpc.1.9.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Castresana C., Garcia-Luque I., Alonso E., Malik V. S., Cashmore A. R. Both positive and negative regulatory elements mediate expression of a photoregulated CAB gene from Nicotiana plumbaginifolia. EMBO J. 1988 Jul;7(7):1929–1936. doi: 10.1002/j.1460-2075.1988.tb03030.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen Z. L., Pan N. S., Beachy R. N. A DNA sequence element that confers seed-specific enhancement to a constitutive promoter. EMBO J. 1988 Feb;7(2):297–302. doi: 10.1002/j.1460-2075.1988.tb02812.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Colot V., Robert L. S., Kavanagh T. A., Bevan M. W., Thompson R. D. Localization of sequences in wheat endosperm protein genes which confer tissue-specific expression in tobacco. EMBO J. 1987;6(12):3559–3564. doi: 10.1002/j.1460-2075.1987.tb02685.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dean C., Jones J., Favreau M., Dunsmuir P., Bedbrook J. Influence of flanking sequences on variability in expression levels of an introduced gene in transgenic tobacco plants. Nucleic Acids Res. 1988 Oct 11;16(19):9267–9283. doi: 10.1093/nar/16.19.9267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jefferson R. A., Burgess S. M., Hirsh D. beta-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8447–8451. doi: 10.1073/pnas.83.22.8447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jofuku K. D., Okamuro J. K., Goldberg R. B. Interaction of an embryo DNA binding protein with a soybean lectin gene upstream region. Nature. 1987 Aug 20;328(6132):734–737. doi: 10.1038/328734a0. [DOI] [PubMed] [Google Scholar]
  11. Kuhlemeier C., Fluhr R., Green P. J., Chua N. H. Sequences in the pea rbcS-3A gene have homology to constitutive mammalian enhancers but function as negative regulatory elements. Genes Dev. 1987 May;1(3):247–255. doi: 10.1101/gad.1.3.247. [DOI] [PubMed] [Google Scholar]
  12. Kuwabara M. D., Sigman D. S. Footprinting DNA-protein complexes in situ following gel retardation assays using 1,10-phenanthroline-copper ion: Escherichia coli RNA polymerase-lac promoter complexes. Biochemistry. 1987 Nov 17;26(23):7234–7238. doi: 10.1021/bi00397a006. [DOI] [PubMed] [Google Scholar]
  13. Maier U. G., Brown J. W., Tologcyzki C., Feix G. Binding of a nuclear factor to a consensus sequence in the 5' flanking region of zein genes from maize. EMBO J. 1987 Jan;6(1):17–22. doi: 10.1002/j.1460-2075.1987.tb04712.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Okamuro J. K., Jofuku K. D., Goldberg R. B. Soybean seed lectin gene and flanking nonseed protein genes are developmentally regulated in transformed tobacco plants. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8240–8244. doi: 10.1073/pnas.83.21.8240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Reiss B., Sprengel R., Will H., Schaller H. A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene. 1984 Oct;30(1-3):211–217. doi: 10.1016/0378-1119(84)90122-7. [DOI] [PubMed] [Google Scholar]
  16. Sanders P. R., Winter J. A., Barnason A. R., Rogers S. G., Fraley R. T. Comparison of cauliflower mosaic virus 35S and nopaline synthase promoters in transgenic plants. Nucleic Acids Res. 1987 Feb 25;15(4):1543–1558. doi: 10.1093/nar/15.4.1543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schernthaner J. P., Matzke M. A., Matzke A. J. Endosperm-specific activity of a zein gene promoter in transgenic tobacco plants. EMBO J. 1988 May;7(5):1249–1255. doi: 10.1002/j.1460-2075.1988.tb02938.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sengupta-Gopalan C., Reichert N. A., Barker R. F., Hall T. C., Kemp J. D. Developmentally regulated expression of the bean beta-phaseolin gene in tobacco seed. Proc Natl Acad Sci U S A. 1985 May;82(10):3320–3324. doi: 10.1073/pnas.82.10.3320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Vonder Haar R. A., Allen R. D., Cohen E. A., Nessler C. L., Thomas T. L. Organization of the sunflower 11S storage protein gene family. Gene. 1988 Dec 30;74(2):433–443. doi: 10.1016/0378-1119(88)90176-x. [DOI] [PubMed] [Google Scholar]
  20. Watt F., Molloy P. L. High mobility group proteins 1 and 2 stimulate binding of a specific transcription factor to the adenovirus major late promoter. Nucleic Acids Res. 1988 Feb 25;16(4):1471–1486. doi: 10.1093/nar/16.4.1471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wright J. M., Dixon G. H. Induction by torsional stress of an altered DNA conformation 5' upstream of the gene for a high mobility group protein from trout and specific binding to flanking sequences by the gene product HMG-T. Biochemistry. 1988 Jan 26;27(2):576–581. doi: 10.1021/bi00402a012. [DOI] [PubMed] [Google Scholar]

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