Skip to main content
SpringerLink
Log in

Anaerobiosis-specific interaction of tobacco nuclear factors with cis-regulatory sequences in the maize GapC4 promoter

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

The promoter of the maize glyceraldehyde-3-phosphate dehydrogenase 4 gene (GapC4) confers strong, specific and ubiquitous anaerobic reporter gene expression in tobacco. To identify factors required for heterologous anaerobic gene expression, 19 progressive 5′ and 3′ promoter deletions were linked to a chimeric GapC4 TATA box-β-glucuronidase (GUS) reporter gene construct and transformed into tobacco. In all transgenic lines aerobic expression values were in the range obtained for negative controls while histochemical GUS assays reveal some weak expression in roots only. Anaerobic induction of about 100-fold to more than 1000-fold above unspecific background is mediated by a region of about 190 bp of the GapC4 promoter. Anaerobic reporter gene induction strongly decreases upon deletion of a 20 bp fragment from −286 to −266 relative to the transcription start point. This fragment harbours putative cis-acting sequences. Electrophoretic mobility shift assays with a 50 bp fragment harbouring these cis sequences reveal a high-mobility complex that is formed with nuclear extracts from aerobic and anaerobic leaf tissue while an additional low-mobility complex is anaerobiosis-specific. The formation of the high-mobility complex requires the sequence GTGGGCCCG. The 50 bp fragment alone confers weak and orientation-dependent anaerobic induction to a GapC4 TATA box-β-glucuronidase (GUS) reporter gene.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. 1988. Current Protocols in Molecular Biology. Greene/Wiley Interscience, New York.

    Google Scholar 

  • Benfey, P.N., Ren, L. and Chua, N.H. 1989. The CaMV 35S enhancer contains at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J. 8: 2195–2202.

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 7: 248–254.

    Google Scholar 

  • Bucher, M., Brander, K.A., Sbicego, S., Mandel, T. and Kuhlemeier, C. 1995. Aerobic fermentation in tobacco pollen. Plant Mol. Biol. 28: 739–750.

    Google Scholar 

  • Bülow, L., Köhler, U., Cerff, R., Hehl, R. and Düring, K. 1999. Induction of the maize GapC4 promoter in transgenic potato under anaerobiosis and in Erwinia carotovora- inoculated tuber tissue. Mol. Plant-Microbe Interact 12: 182–188.

    Google Scholar 

  • de Vetten, N.C. and Ferl, R.J. 1995. Characterization of a maize G-box binding factor that is induced by hypoxia. Plant J. 7: 589–601.

    Google Scholar 

  • Dolferus, R., Jacobs, M., Peacock, W.J. and Dennis, E.S. 1994. Differential interactions of promoter elements in stress responses of the Arabidopsis Adh gene. Plant Physiol. 105: 1075–1087.

    Google Scholar 

  • Drew, M.C. 1997. Oxygen deficiency and root metabolism: injury and acclimation under hypoxia and anoxia. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 223–250.

    Google Scholar 

  • Ellis, J.G., Llewellyn, D.J., Dennis, E.S. and Peacock, W.J. 1987. Maize Adh-1 promoter sequences control anaerobic regulation: addition of upstream promoter elements from constitutive genes is necessary for expression in tobacco. EMBO J. 6: 11–16.

    Google Scholar 

  • Ferl, R.J. 1990. ARF-B2: a protein complex that specifically binds to part of the anaerobic response element of maize Adh1. Plant Physiol. 93: 1094–1101.

    Google Scholar 

  • Ferl, R.J. 1996. 14-3-3 proteins and signal transduction. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47: 49–73.

    Google Scholar 

  • Gatz, C. 1997. Chemical control of gene expression. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 89–108.

    Google Scholar 

  • Hehl, R. and Baker, B. 1989. Induced transposition of Ds by a stable Ac in crosses of transgenic tobacco plants. Mol. Gen. Genet 217: 53–59.

    Google Scholar 

  • Hoeren, F.U., Dolferus, R., Wu, Y., Peacock, W.J. and Dennis, E.S. 1998. Evidence for a role for AtMYB2 in the induction of the Arabidopsis alcohol dehydrogenase gene (ADH1) by low oxygen. Genetics 149: 479–490.

    Google Scholar 

  • Honma, M.A., Baker, B.J. and Waddell, C.S. 1993. High-frequency transposition of Ds-ALS in Arabidopsis. Proc. Natl. Acad. Sci. USA 90: 6242–6246.

    Google Scholar 

  • Hood, E.E., Helmer, G.L., Fraley, R.T. and Chilton, M.D. 1986. The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. J. Bacteriol. 168: 1291–301.

    Google Scholar 

  • Horsch, R.B., Fry, J.E., Hoffmann, N.L., Eichholtz, D., Rogers, S.G. and Fraley, R.T. 1985. A simple and general method for transferring genes into plants. Science 227: 1229–1231.

    Google Scholar 

  • Jefferson, R.A. 1987. Assaying chimaric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5: 387–405.

    Google Scholar 

  • Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. 1987. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907.

    Google Scholar 

  • Köhler, U., Liaud, M.-F., Mendel, R.R., Cerff, R. and Hehl, R. 1995. The maize GapC4 promoter confers anaerobic reporter gene expression and shows homology to the maize anthocyanin regulatory locus C1. Plant Mol. Biol. 29: 1293–1298.

    Google Scholar 

  • Köhler, U., Donath, M., Mendel, R.R., Cerff, R. and Hehl, R. 1996a. Intron-specific stimulation of anaerobic gene expression and splicing efficiency in maize cells. Mol. Gen. Genet. 251: 252–258.

    Google Scholar 

  • Köhler, U., Mendel, R.R., Cerff, R. and Hehl, R. 1996b. A promoter for strong and ubiquitous anaerobic gene expression in tobacco. Plant J. 10: 175–183.

    Google Scholar 

  • Kyozuka, J., Fujimoto, H., Izawa, T. and Shimamoto, K. 1991. Anaerobic induction and tissue-specific expression of maize Adh1 promoter in transgenic rice plants and their progeny. Mol. Gen. Genet. 228: 40–48.

    Google Scholar 

  • Lu, G., DeLisle, A.J., de Vetten, N.C. and Ferl, R.J. 1992. Brain proteins in plants: an Arabidopsis homolog to neurotransmitter pathway activators is part of a DNA binding complex. Proc. Natl. Acad. Sci. USA 89: 11490–11494.

    Google Scholar 

  • Lu, G., Sehnke, P.C. and Ferl, R.J. 1994. Phosphorylation and calcium binding properties of an Arabidopsis GF14 brain protein homolog. Plant Cell 6: 501–510.

    Google Scholar 

  • Manjunath, S. and Sachs, M.M. 1997. Molecular characterization and promoter analysis of the maize cytosolic glyceraldehyde 3-phosphate dehydrogenase gene family and its expression during anoxia. Plant Mol. Biol. 33: 97–112.

    Google Scholar 

  • Nagel, R., Elliott, A., Masel, A., Birch, R.G. and Manners, J.M. 1990. Electroporation of binary Ti plasmid vector into Agrobacterium tumefaciens and Agrobacterium rhizogenes. FEMS 67: 325–328.

    Google Scholar 

  • Olive, M.R., Walker, J.C., Singh, K., Dennis, E.S. and Peacock, W.J. 1990. Functional properties of the anaerobic responsive element of the maize Adh1 gene. Plant Mol. Biol. 15: 593–604.

    Google Scholar 

  • Olive, M.R., Peacock, W.J. and Dennis, E.S. 1991. The anerobic responsive element contains two GC-rich sequences essential for binding a nuclear protein and hypoxic activation of the maize Adh1 promoter. Nucl. Acids Res. 19: 7053–7060.

    Google Scholar 

  • Olszewski, N.E., Martin, F.B. and Ausubel, F.M. 1988. Specialized binary vector for plant transformation: expression of the Arabidopsis thaliana AHAS gene in Nicotiana tabacum. Nucl. Acids Res. 16: 10765–10782.

    Google Scholar 

  • Parker, C.S. and Topol, J. 1984. A Drosophila polymerase II transcription factor contains a promotor region specific DNA binding activity. Cell 36: 357–369.

    Google Scholar 

  • Paul, A.L. and Ferl, R.J. 1991. In vivo footprinting reveals unique cis-elements and different modes of hypoxic induction in maize Adh1 and Adh2. Plant Cell 3: 159–168.

    Google Scholar 

  • Sachs, M.M., Freeling, M. and Okimoto, R. 1980. The anaerobic proteins of maize. Cell 20: 761–767.

    Google Scholar 

  • Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  • Sanger, M., Daubert, S. and Goodman, R.M. 1990. Characteristics of a strong promoter from figwort mosaic virus: comparison with the analogous 35S promoter from cauliflower mosaic virus and the regulated mannopine synthase promoter. Plant Mol. Biol. 14: 433–443.

    Google Scholar 

  • Subbaiah, C.C., Bush, D.S. and Sachs, M.M. 1994a. Elevation of cytosolic calcium precedes anoxic gene expression in maize suspension-cultured cells. Plant Cell 6: 1747–1762.

    Google Scholar 

  • Subbaiah, C.C., Zhang, J. and Sachs, M.M. 1994b. Involvement of intracellular calcium in anaerobic gene expression and survival of maize seedlings. Plant Physiol. 105: 369–376.

    Google Scholar 

  • Walker, J.C., Howard, E.A., Dennis, E.S. and Peacock, W.J. 1987. DNA sequences required for anaerobic expression of the maize alcohol dehydrogenase 1 gene. Proc. Natl. Acad. Sci. USA 84: 6624–6628.

    Google Scholar 

  • Watson, J.C. and Thompson, W.F. 1986. Purification and restriction endonucleases analysis of plant nuclear DNA. Meth. Enzymol. 118: 57–75.

    Google Scholar 

  • Williams, M.E., Foster, R. and Chua, N.-H. 1992. Sequences flanking the hexameric G-box core CACGTG affect the specificity of protein binding. Plant Cell 4: 485–496.

    Google Scholar 

  • Yang, Y., Kwon, H.-B., Peng, H.-P. and Shih, M.-C. 1993. Stress responses and metabolic regulation of glyceraldehyde-3-phosphate dehydrogenase genes in Arabidopsis. Plant Physiol. 101: 209–216.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Genetics, Technical University of Braunschweig, Spielmannstrasse 7, 38106, Braunschweig, Germany

    Robert Geffers, Rüdiger Cerff & Reinhard Hehl

Authors
  1. Robert Geffers
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rüdiger Cerff
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Reinhard Hehl
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Geffers, R., Cerff, R. & Hehl, R. Anaerobiosis-specific interaction of tobacco nuclear factors with cis-regulatory sequences in the maize GapC4 promoter. Plant Mol Biol 43, 11–21 (2000). https://doi.org/10.1023/A:1006419232075

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006419232075

Search

Navigation