Skip to main content
PMC home page
The Plant Cell logoLink to The Plant Cell
. 1994 Feb;6(2):237–249. doi: 10.1105/tpc.6.2.237

The transcriptional activator Opaque2 recognizes two different target sequences in the 22-kD-like alpha-prolamin genes.

J A Yunes 1, G Cord Neto 1, M J da Silva 1, A Leite 1, L M Ottoboni 1, P Arruda 1
PMCID: PMC160430  PMID: 8148647

Abstract

The maize Opaque2 (O2) protein is a "leucine zipper" DNA binding factor that interacts with the sequence TCCACGTAGA in the promoters of the 22-kD alpha-zein genes and activates its transcription. A completely different consensus sequence (GATGAPyPuTGPu) identified in b-32, a gene that encodes an abundant albumin that is also under control of the O2 locus, can also be bound by the O2 protein. We showed that the gene encoding the 22-kD-like alpha-coixin, the alpha-prolamin of the maize-related grass Coix, can also be transactivated by the O2 protein. A binding assay in vitro and footprint analysis demonstrated that the GACATGTC sequence of the alpha-coixin promoter can be recognized and protected by the maize O2 protein. Employing transient expression experiments in immature maize endosperm and tobacco mesophyll protoplasts, we demonstrated that the O2 protein can activate expression of the beta-glucuronidase reporter gene placed under the control of the 22-kD-like alpha-coixin promoter. We also demonstrated that a 22-kD-like alpha-coixin pseudogene promoter is transactivated by the maize O2 protein.

Full Text

The Full Text of this article is available as a PDF (3.1 MB).

Selected References

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

  1. Armstrong G. A., Weisshaar B., Hahlbrock K. Homodimeric and heterodimeric leucine zipper proteins and nuclear factors from parsley recognize diverse promoter elements with ACGT cores. Plant Cell. 1992 May;4(5):525–537. doi: 10.1105/tpc.4.5.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Burr B., Burr F. A., St John T. P., Thomas M., Davis R. W. Zein storage protein gene family of maize. An assessment of heterogeneity with cloned messenger RNA sequences. J Mol Biol. 1982 Jan 5;154(1):33–49. doi: 10.1016/0022-2836(82)90415-6. [DOI] [PubMed] [Google Scholar]
  4. Burr F. A., Burr B. Three mutations in Zea mays affecting zein accumulation: a comparison of zein polypeptides, in vitro synthesis and processing, mRNA levels, and genomic organization. J Cell Biol. 1982 Jul;94(1):201–206. doi: 10.1083/jcb.94.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Entwistle J., Knudsen S., Müller M., Cameron-Mills V. Amber codon suppression: the in vivo and in vitro analysis of two C-hordein genes from barley. Plant Mol Biol. 1991 Dec;17(6):1217–1231. doi: 10.1007/BF00028737. [DOI] [PubMed] [Google Scholar]
  6. Esen A. Separation of alcohol-soluble proteins (zeins) from maize into three fractions by differential solubility. Plant Physiol. 1986 Mar;80(3):623–627. doi: 10.1104/pp.80.3.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. A comparison of eukaryotic viral 5'-leader sequences as enhancers of mRNA expression in vivo. Nucleic Acids Res. 1987 Nov 11;15(21):8693–8711. doi: 10.1093/nar/15.21.8693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hagen G., Rubenstein I. Complex organization of zein genes in maize. Gene. 1981 Apr;13(3):239–249. doi: 10.1016/0378-1119(81)90029-9. [DOI] [PubMed] [Google Scholar]
  9. Hammond-Kosack M. C., Holdsworth M. J., Bevan M. W. In vivo footprinting of a low molecular weight glutenin gene (LMWG-1D1) in wheat endosperm. EMBO J. 1993 Feb;12(2):545–554. doi: 10.1002/j.1460-2075.1993.tb05686.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hill D. E., Hope I. A., Macke J. P., Struhl K. Saturation mutagenesis of the yeast his3 regulatory site: requirements for transcriptional induction and for binding by GCN4 activator protein. Science. 1986 Oct 24;234(4775):451–457. doi: 10.1126/science.3532321. [DOI] [PubMed] [Google Scholar]
  11. Izawa T., Foster R., Chua N. H. Plant bZIP protein DNA binding specificity. J Mol Biol. 1993 Apr 20;230(4):1131–1144. doi: 10.1006/jmbi.1993.1230. [DOI] [PubMed] [Google Scholar]
  12. Kodrzycki R., Boston R. S., Larkins B. A. The opaque-2 mutation of maize differentially reduces zein gene transcription. Plant Cell. 1989 Jan;1(1):105–114. doi: 10.1105/tpc.1.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leite A., Ottoboni L. M., Targon M. L., Silva M. J., Turcinelli S. R., Arruda P. Phylogenetic relationship of zeins and coixins as determined by immunological cross-reactivity and Southern blot analysis. Plant Mol Biol. 1990 May;14(5):743–751. doi: 10.1007/BF00016507. [DOI] [PubMed] [Google Scholar]
  14. Liu C. N., Rubenstein I. Genomic organization of an alpha-zein gene cluster in maize. Mol Gen Genet. 1992 Jan;231(2):304–312. doi: 10.1007/BF00279804. [DOI] [PubMed] [Google Scholar]
  15. Lohmer S., Maddaloni M., Motto M., Di Fonzo N., Hartings H., Salamini F., Thompson R. D. The maize regulatory locus Opaque-2 encodes a DNA-binding protein which activates the transcription of the b-32 gene. EMBO J. 1991 Mar;10(3):617–624. doi: 10.1002/j.1460-2075.1991.tb07989.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Oeda K., Salinas J., Chua N. H. A tobacco bZip transcription activator (TAF-1) binds to a G-box-like motif conserved in plant genes. EMBO J. 1991 Jul;10(7):1793–1802. doi: 10.1002/j.1460-2075.1991.tb07704.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oliphant A. R., Brandl C. J., Struhl K. Defining the sequence specificity of DNA-binding proteins by selecting binding sites from random-sequence oligonucleotides: analysis of yeast GCN4 protein. Mol Cell Biol. 1989 Jul;9(7):2944–2949. doi: 10.1128/mcb.9.7.2944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ottoboni L. M., Leite A., Yunes J. A., Targon M. L., de Souza Filho G. A., Arruda P. Sequence analysis of 22 kDa-like alpha-coixin genes and their comparison with homologous zein and kafirin genes reveals highly conserved protein structure and regulatory elements. Plant Mol Biol. 1993 Mar;21(5):765–778. doi: 10.1007/BF00027110. [DOI] [PubMed] [Google Scholar]
  19. Pysh L. D., Aukerman M. J., Schmidt R. J. OHP1: a maize basic domain/leucine zipper protein that interacts with opaque2. Plant Cell. 1993 Feb;5(2):227–236. doi: 10.1105/tpc.5.2.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schmidt R. J., Burr F. A., Aukerman M. J., Burr B. Maize regulatory gene opaque-2 encodes a protein with a "leucine-zipper" motif that binds to zein DNA. Proc Natl Acad Sci U S A. 1990 Jan;87(1):46–50. doi: 10.1073/pnas.87.1.46. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schmidt R. J., Burr F. A., Burr B. Transposon tagging and molecular analysis of the maize regulatory locus opaque-2. Science. 1987 Nov 13;238(4829):960–963. doi: 10.1126/science.2823388. [DOI] [PubMed] [Google Scholar]
  22. Schmidt R. J., Ketudat M., Aukerman M. J., Hoschek G. Opaque-2 is a transcriptional activator that recognizes a specific target site in 22-kD zein genes. Plant Cell. 1992 Jun;4(6):689–700. doi: 10.1105/tpc.4.6.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Spena A., Viotti A., Pirrotta V. A homologous repetitive block structure underlies the heterogeneity of heavy and light chain zein genes. EMBO J. 1982;1(12):1589–1594. doi: 10.1002/j.1460-2075.1982.tb01360.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Thompson G. A., Siemieniak D. R., Sieu L. C., Slightom J. L., Larkins B. A. Sequence analysis of linked maize 22 kDa alpha-zein genes. Plant Mol Biol. 1992 Feb;18(4):827–833. doi: 10.1007/BF00020030. [DOI] [PubMed] [Google Scholar]
  25. Töpfer R., Matzeit V., Gronenborn B., Schell J., Steinbiss H. H. A set of plant expression vectors for transcriptional and translational fusions. Nucleic Acids Res. 1987 Jul 24;15(14):5890–5890. doi: 10.1093/nar/15.14.5890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wandelt C., Feix G. Sequence of a 21 kd zein gene from maize containing an in-frame stop codon. Nucleic Acids Res. 1989 Mar 25;17(6):2354–2354. doi: 10.1093/nar/17.6.2354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ye G. N., Daniell H., Sanford J. C. Optimization of delivery of foreign DNA into higher-plant chloroplasts. Plant Mol Biol. 1990 Dec;15(6):809–819. doi: 10.1007/BF00039421. [DOI] [PubMed] [Google Scholar]

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

RESOURCES