Skip to main content
PMC home page
The Plant Cell logoLink to The Plant Cell
. 1990 Mar;2(3):207–214. doi: 10.1105/tpc.2.3.207

In vivo and in vitro characterization of protein interactions with the dyad G-box of the Arabidopsis Adh gene.

W L McKendree 1, A L Paul 1, A J DeLisle 1, R J Ferl 1
PMCID: PMC159877  PMID: 2152112

Abstract

Expression of the alcohol dehydrogenase (Adh) and ribulose-1,5-bisphosphate carboxylase small subunit (RbcS) genes of higher plants is cell-type-specific and environmentally inducible. However, the tissues in which these two genes are expressed, their modes of induction, and their protein functions are quite distinct. Adh is expressed in non-green tissue, induced by anaerobiosis, and repressed in leaves. RbcS is only expressed in green tissue. An 8-base pair G-box element (5'-CCACGTGG-3') is associated with light-induced expression of RbcS and chalcone synthase. The same sequence is also present in the 5'-flanking region of Arabidopsis thaliana Adh, and this sequence is associated with a trans-acting factor in vivo. We report here that in vitro Adh G-box binding activity is present in crude whole cell extracts of both cell culture and leaves of Arabidopsis. The authenticity of in vitro Adh G-box binding is supported by in vivo and in vitro dimethylsulfate footprinting. A clear in vivo Adh G-box footprint occurs in cell cultures, but comparable in vivo binding to the Adh G-box does not occur in leaves. Therefore, there does not appear to be a direct correlation between the presence of the G-box factor in a tissue and its binding to the Adh G-box.

Full Text

The Full Text of this article is available as a PDF (2.0 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chang C., Meyerowitz E. M. Molecular cloning and DNA sequence of the Arabidopsis thaliana alcohol dehydrogenase gene. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1408–1412. doi: 10.1073/pnas.83.5.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chodosh L. A., Buratowski S., Sharp P. A. A yeast protein possesses the DNA-binding properties of the adenovirus major late transcription factor. Mol Cell Biol. 1989 Feb;9(2):820–822. doi: 10.1128/mcb.9.2.820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dennis E. S., Gerlach W. L., Pryor A. J., Bennetzen J. L., Inglis A., Llewellyn D., Sachs M. M., Ferl R. J., Peacock W. J. Molecular analysis of the alcohol dehydrogenase (Adh1) gene of maize. Nucleic Acids Res. 1984 May 11;12(9):3983–4000. doi: 10.1093/nar/12.9.3983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dennis E. S., Sachs M. M., Gerlach W. L., Finnegan E. J., Peacock W. J. Molecular analysis of the alcohol dehydrogenase 2 (Adh2) gene of maize. Nucleic Acids Res. 1985 Feb 11;13(3):727–743. doi: 10.1093/nar/13.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dynan W. S., Tjian R. Control of eukaryotic messenger RNA synthesis by sequence-specific DNA-binding proteins. 1985 Aug 29-Sep 4Nature. 316(6031):774–778. doi: 10.1038/316774a0. [DOI] [PubMed] [Google Scholar]
  7. Ferl R. J., Brennan M. D., Schwartz D. In vitro translation of maize ADH: evidence for the anaerobic induction of mRNA. Biochem Genet. 1980 Aug;18(7-8):681–691. doi: 10.1007/BF00484585. [DOI] [PubMed] [Google Scholar]
  8. Ferl R. J., Nick H. S. In vivo detection of regulatory factor binding sites in the 5' flanking region of maize Adh1. J Biol Chem. 1987 Jun 15;262(17):7947–7950. [PubMed] [Google Scholar]
  9. Finkelstein R. R., Tenbarge K. M., Shumway J. E., Crouch M. L. Role of ABA in Maturation of Rapeseed Embryos. Plant Physiol. 1985 Jul;78(3):630–636. doi: 10.1104/pp.78.3.630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Gerlach W. L., Pryor A. J., Dennis E. S., Ferl R. J., Sachs M. M., Peacock W. J. cDNA cloning and induction of the alcohol dehydrogenase gene (Adh1) of maize. Proc Natl Acad Sci U S A. 1982 May;79(9):2981–2985. doi: 10.1073/pnas.79.9.2981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Giuliano G., Pichersky E., Malik V. S., Timko M. P., Scolnik P. A., Cashmore A. R. An evolutionarily conserved protein binding sequence upstream of a plant light-regulated gene. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7089–7093. doi: 10.1073/pnas.85.19.7089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Kuhlemeier C., Strittmatter G., Ward K., Chua N. H. The Pea rbcS-3A Promoter Mediates Light Responsiveness but not Organ Specificity. Plant Cell. 1989 Apr;1(4):471–478. doi: 10.1105/tpc.1.4.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lee L., Fenoll C., Bennetzen J. L. Construction and Homologous Expression of a Maize Adh1 Based NcoI Cassette Vector. Plant Physiol. 1987 Oct;85(2):327–330. doi: 10.1104/pp.85.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nagy F., Morelli G., Fraley R. T., Rogers S. G., Chua N. H. Photoregulated expression of a pea rbcS gene in leaves of transgenic plants. EMBO J. 1985 Dec 1;4(12):3063–3068. doi: 10.1002/j.1460-2075.1985.tb04046.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rowland L. J., Strommer J. N. Anaerobic treatment of maize roots affects transcription of Adh1 and transcript stability. Mol Cell Biol. 1986 Oct;6(10):3368–3372. doi: 10.1128/mcb.6.10.3368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sachs M. M., Freeling M., Okimoto R. The anaerobic proteins of maize. Cell. 1980 Jul;20(3):761–767. doi: 10.1016/0092-8674(80)90322-0. [DOI] [PubMed] [Google Scholar]
  19. Schulze-Lefert P., Dangl J. L., Becker-André M., Hahlbrock K., Schulz W. Inducible in vivo DNA footprints define sequences necessary for UV light activation of the parsley chalcone synthase gene. EMBO J. 1989 Mar;8(3):651–656. doi: 10.1002/j.1460-2075.1989.tb03422.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shuey D. J., Parker C. S. Binding of Drosophila heat-shock gene transcription factor to the hsp 70 promoter. Evidence for symmetric and dynamic interactions. J Biol Chem. 1986 Jun 15;261(17):7934–7940. [PubMed] [Google Scholar]
  21. Sugita M., Gruissem W. Developmental, organ-specific, and light-dependent expression of the tomato ribulose-1,5-bisphosphate carboxylase small subunit gene family. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7104–7108. doi: 10.1073/pnas.84.20.7104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Treisman R. Identification of a protein-binding site that mediates transcriptional response of the c-fos gene to serum factors. Cell. 1986 Aug 15;46(4):567–574. doi: 10.1016/0092-8674(86)90882-2. [DOI] [PubMed] [Google Scholar]
  23. Walker J. C., Howard E. A., Dennis E. S., Peacock W. J. DNA sequences required for anaerobic expression of the maize alcohol dehydrogenase 1 gene. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6624–6628. doi: 10.1073/pnas.84.19.6624. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES