Skip to main content
SpringerLink
Log in

Expression of the auxin-regulated parA gene in transgenic tobacco and nuclear localization of its gene products

  • Published:
Planta Aims and scope Submit manuscript

Abstract

An auxin-regulated gene, parA, comprises a gene family consisting of a handful genes which respond to various signals. Although Droog et al. (Plant Mol. Biol, 1993, 21, 965–972) postulated that the parA-related genes belong to the family of a cytoplasmic enzyme, glutathione S-transferase (GST), we detected a low level of GST activity in the parA products, whose value was below 1/30 of that of parB products encoding tobacco (Nicotiana tabacum L.) GST. Immunofluorescence studies using an antibody against parA protein revealed that the subcellular location of parA protein is the nucleus in cultured tobacco mesophyll protoplasts, while conventional GSTs' including the parB product were primarily located in the cytoplasm. Confocal laser scanning microscopy of tobacco BY-2 cells showed that the parA product was confined to the nucleus, but was excluded from the nucleolus. In addition, exon/intron organization of the parA family was appreciably different from that of conventional GSTs including parB. Furthermore, the parA protein is much more similar to a 24-kDa protein of Escherichia coli that is reported to bind to RNA polymerase. These different characteristics of parA compared with to the conventional GSTs, indicate that parA protein would have distinct functions, such as involvement in transcription, rather than functioning as a conventional GST. Transgenic tobacco plants that carried the parA promoter fused to a β-glucuronidase gene were used to show that the parA gene is tissue-specific and also under developmental control.

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

Abbreviations

GST:

glutathione S-transferase

GUS:

β-glucuronidase

MBP:

maltose-binding protein

IPTG:

isopropyl β-d-thiogalactopyranoside

2,4-D:

2,4-dichlorophenoxyacetic acid

DAPI:

4', 6-diamidino-2-phenylindole

References

  • Allison, L.A., Mayle, M., Shales, M., Ingres, J. (1985) Extensive homology among the largest subunits of eukaryotic and prokaryotic RNA polymerase. Cell 42, 589–610

    Google Scholar 

  • Amann, E., Broslus, J. (1985) ‘ATG vectors’ for regulated high-level expression of cloned genes in Escherichia coli. Gene 40, 183

    Google Scholar 

  • Biggs, J., Searles, L.L., Greenleaf, A.L. (1985) Structure of the eukaryotic polymerase II. Cell 42, 611–621

    Google Scholar 

  • Coschigano, P.W., Magasanik, B. (1991) The URE2 gene product of Saccharomyces cerevisiae plays an important role in the cellular response to the nitrogen source has homology to glutathione S-transferases. Mol. Cell. Biol. 11, 822–832

    Google Scholar 

  • Czarnecka, E., Nagao, R.T., Key, J.L., Gurley, W.B. (1988) Characterization of Gmhsp 26-A, a stress gene encoding a divergent heat shock protein of soybean: heavy-metal-induced inhibition of intron processing. Mol. Cell. Biol. 8, 1113–1122

    Google Scholar 

  • Dingwall, C., Laskey, R.A. (1991) Nuclear targeting sequences-a consensus? Trends Biochem. Sci. 16, 478–481

    Google Scholar 

  • Dominov, J.A., Stenzler, L., Lee, S., Schwartz, J.J., Leinser, S., Howell, S.S. (1992) Cytokinins and auxins control the expression of a gene in Nicotiana plumbaginifolia cells by feedback regulation. Plant Cell 4, 451–461

    Google Scholar 

  • Droog, F.N.J., Hooykaas, P.J.J., Libbenga, K.R., van der Zaal, E.J. (1993) Proteins encoded by an auxin-regulated gene family of tobacco share limited but significant homology with glutathione S-transferase and one member indeed shows in vitro GST activity. Plant Mol. Biol. 21, 965–972

    Google Scholar 

  • Dudler, R., Hertig, C., Rebman, G., Bull, J., Mauch, F. (1991) A pathogen-induced wheat gene encodes a protein homologous to glutathione S-transferase. Mol. Plant-Microbe Interact. 4, 14–18

    Google Scholar 

  • Grove, G., Zarlengo, R.P., Timmerman, K.P., Li, N.-P., Tam, M.F., Tu, C.-P. (1988) Characterization and heterospecific expression of cDNA clones of genes in the maize GSH S-transferase mutigene family. Nucleic Acids Res. 16, 425–438

    Google Scholar 

  • Guan, C., Li, P., Riggs, P.D., Inoue, H. (1987) Vectors that facilitate the expression and purification of foreign peptides in Escherichia coli by fusion to maltose-binding protein. Gene 67, 21–30

    Google Scholar 

  • Guilfoyle, T.J. (1986) Auxin-regulated gene expression in higher plants. CRC Crit. Rev. Plant Sci. 4, 247–276

    Google Scholar 

  • Hagen, G., Martin, G., Li, Y., Guilfoyle, T.J. (1991) Auxin-induced expression of the soybean GH3 promoter in transgenic tobacco plants. Plant Mol. Biol. 17, 567–579

    Google Scholar 

  • Hall, M.N., Hereford, L., Herskowitz, I. (1984) Targeting of E. coli β-galactosidase to the nucleus in yeast. Cell 36, 1057–1065

    Google Scholar 

  • Hasezawa, S., Nagata, T. (1991) Dynamic organization of plant microtubules at the three distinct transition points during the cell cycle progression of synchronized tobacco BY-2 cells. Bot. Acta 104, 206–211

    Google Scholar 

  • Ishida, S., Takahashi, Y., Nagata, T. (1993) Isolation of cDNA of an auxin-regulated gene encoding a G protein β subunit-like protein from tobacco BY-2 cells. Proc. Natl. Acad. Sci. USA 90, 11152–11156

    Google Scholar 

  • Ishihama, A., Saitoh, T. (1979) Subunits of RNA polymerase in function and structure. IX. Regulation of RNA polymerase activity by stringent starvation protein (ssp). J. Mol. Biol. 129, 517–530

    Google Scholar 

  • Jefferson, R.A. (1987) Assaying chimeric genes in plants. The GUS gene fusion system. Plant Mol. Biol. Rep. 5, 387–405

    Google Scholar 

  • Kaldeson, D., Richardson, W.D., Markham, F.A., Smith, A.E. (1984) Sequence requirements for nuclear location of simian virus 40 large T antigen. Nature 311, 33–38

    Google Scholar 

  • Li, Y., Hagen, G., Guilfoyle, T.J. (1991) An auxin-responsive promoter is differentially induced by auxin gradients during tropism. Plant Cell 3, 1167–1175

    Google Scholar 

  • Marc, J., Gunning, B.E.S. (1988) Monoclonal antibodies to a fern spermatozoid detect novel components of the mitotic and cytokinetic apparatus in higher plant cells. Protoplasma 142, 15–24

    Google Scholar 

  • Maurel, C., Brevet, J., Barbier-Brygoo, H., Guern, J., Tempé, J. (1990) Auxin regulates the promoter of the root-inducing rolB gene of Agrobacterium rhizogenes in transgenic tobacco. Mol. Gen. Genet. 223, 58–64

    Google Scholar 

  • Meyer, R.C., Jr., Goldsbrough, P.B., Wodson, W.R. (1991) An ethylene-responsive flower senescence-related gene from carnation encodes a protein homologous to glutathione S-transferase. Plant Mol. Biol. 17, 277–281

    Google Scholar 

  • Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497

    Google Scholar 

  • Nagata, T., Takebe, T. (1970) Cell wall regeneration and cell division in isolated tobacco mesophyll protoplasts. Planta 92, 201–208

    Google Scholar 

  • Nash, J., Luehrsen, K.R., Walbot, V. (1990) Bronze-2 gene of maize: Reconstruction of a wild-type allele and analysis of transcription and splicing. Plant Cell 2, 1039–1049

    Google Scholar 

  • Raikhel, N. (1992) Nuclear targeting in plants. Plant Physiol. 100, 1627–1632

    Google Scholar 

  • Serizawa, H., Fukuda, R. (1987) Structure of the gene for the stringent starvation protein of Escherichia coli. Nucleic Acids Res. 15, 1153–1163

    Google Scholar 

  • Shah, D.M., Hironaka, C.M., Wiegnand, R.C., Harding, E.I., Krivi, G.G., Tiemeier, D.C. (1986) Structural analysis of a maize gene coding for glutathione S-transferase involved in herbicide detoxification. Plant Mol. Biol. 6, 203–211

    Google Scholar 

  • Simon, M.I., Strathmann, M.P., Gautam, N. (1991) Diversity of G proteins in signal transduction Science 252, 802–808

    Google Scholar 

  • Studier, F.W., Rosenberg, A.H., Dunn, J.J., Dubendorff, J.W. (1990) Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 185, 60–89

    Google Scholar 

  • Sweetser, D., Monet, M., Young, R.A. (1987) Prokaryotic and eukaryotic RNA polymerases have homologous core subunits. Proc. Natl. Acad. Sci. USA 84, 1192–1196

    Google Scholar 

  • Takahashi, Y., Kuroda, H., Tanaka, T., Machida, Y., Takebe, I., Nagata, T. (1989) Isolation of an auxin-regulated gene cDNA expressed during the transition from G0 to S phase in tobacco mesophyll protoplasts. Proc. Natl. Acad. Sci. USA 86, 9279–9283

    Google Scholar 

  • Takahashi, Y., Niwa, Y., Machida, Y., Nagata, T. (1990) Location of the cis-acting auxin-responsive region in the promoter of the par gene from tobacco mesophyll protoplasts. Proc. Natl. Acad. Sci. USA 87, 8013–8016

    Google Scholar 

  • Takahashi, Y., Kusaba, M., Hiraoka, Y., Nagata, T. (1991) Characterization of the auxin-regulated par gene from tobacco mesophyll protoplasts. Plant J. 1, 327–332

    Google Scholar 

  • Takahashi, Y., Nagata, T. (1992a) parB: An auxin-regulated gene encoding glutathione S-transferase. Proc. Natl. Acad. Sci. USA 89, 56–59

    Google Scholar 

  • Takahashi, Y., Nagata, T. (1992b) Differential expression of an auxin-regulated gene, parC, and a novel related gene, C-7, from tobacco mesophyll protoplasts in response to external stimuli and in plant tissues. Plant Cell Physiol. 33, 779–787

    Google Scholar 

  • Taylor, J.L., Fritzemeier, K.-H., Hauser, I., Kombrink, E., Rohwer, F., Schröder, M., Strittmatter, G., Hahlbrock, K. (1990) Structural analysis and activation by fungal infection of a gene encoding a pathogenesis-related protein in potato. Mol. Plant-Microbe Interact. 3, 72–77

    Google Scholar 

  • Tomarev, S.I., Zinovieva, R.D. (1988) Squid major lens polypeptides are homologous to glutathione S-transferase. Nature 336, 86–88

    Google Scholar 

  • van der Zaal, E.J., Droog, F.N.J., Hensgens, L.A.M., Hoge, J.H.C., Schilperoort, R.A., Libbenga, K.R. (1991) Promoters of auxin-induced genes from tobacco can lead to auxin-inducible and root tip-specific expression. Plant Mol. Biol. 16, 983–998

    Google Scholar 

  • Wareing, P.F., Philips, I.D.J. (1981) Growth and differentiation in plants, 3rd edn. Pergamon, Oxford

    Google Scholar 

  • Went, F.W., Thimann, K.V. (1937) Photohormones, MacMillan, New York

    Google Scholar 

  • Wyatt, R.E., Ainley, W.M., Nagao, R.T., Conner, T.W., Key, J.L. (1993) Expression of the Arabidopsis AtAux3–11 auxin-responsive gene in transgenic plants. Plant Mol. Biol. 22, 731–749

    Google Scholar 

  • Yasuda, T., Kuroiwa, T., Nagata, T. (1988) Preferential synthesis of plastid DNA and increased replication of plastids of cultured tobacco cells following medium renewal. Planta 174, 235–241

    Google Scholar 

  • Zettl, R., Schell, J., Palme, K. Photoaffinity labeling of Arabidopsis thaliana plasma membrane vesicles by 5-azido-[7-3H]indole-3-cetic acid: Identification of a glutathione S-transferase (1994) Proc. Natl. Acad. Sci. USA 91, 689–693

    Google Scholar 

  • Zhou, J., Goldsbrough, P.B. (1993) An Arabidopsis gene with homology to glutathione S-transferase is regulated by ethylene. Plant Mol. Biol. 22, 517–523

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Science, Graduate School of Science, University of Tokyo, Hongo, Bunkyo-ku, 113, Tokyo, Japan

    Yohsuke Takahashi, Seiichiro Hasezawa, Makoto Kusaba & Toshiyuki Nagata

Authors
  1. Yohsuke Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Seiichiro Hasezawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Makoto Kusaba
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Toshiyuki Nagata
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Confocal laser scanning micrograph was taken by the courtesy of Leica Ltd. (Japan). We wish to thank Dr. Dennis Francis (University of Wales, Cardiff, UK) for his critical reading of the manuscript. This study was supported in part by grants from the Ministry of Education, Culture and Science of Japan to T.N. and Y.T. and from the Agency of Science and Technologies of Japan to Y.T.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Takahashi, Y., Hasezawa, S., Kusaba, M. et al. Expression of the auxin-regulated parA gene in transgenic tobacco and nuclear localization of its gene products. Planta 196, 111–117 (1995). https://doi.org/10.1007/BF00193224

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00193224

Key words

Search

Navigation