Skip to main content
SpringerLink
Log in

Specific transcription and reinitiation of 2,4-D-induced genes in tobacco nuclei

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

In order to study the effects oftrans-acting factors oncis-acting elements in plant genes anin vitro reinitiating nuclear transcription system is needed. Here we report that run-on experiments with nuclei isolated from 2,4-D-treated auxin-starved early-stationary-phase cells of tobacco clearly show reinitiation of transcription of specific 2,4-D-induced genes. Using γ-thio nucleotides and [α-32P]-UTP we were able to demonstrate the presence of reinitiated labelled specific RNAs after isolation on a mercury-agarose affinity column. Addition of heparin as an inhibitor blocked this reinitiation. In a primer extension assay we found that the new transcripts initiate at approximately the same site as usedin vivo.

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

2,4-D:

2,4-dichlorophenoxyacetic acid

Tris:

2-amino-2-(hydroxymethyl)-1,3-propanediol

BSA:

bovine serum albumin

SDS:

sodium dodecyl sulphate ME, mercaptoethanol

References

  1. BenzJr EW, Wydro RM, Nadal-Ginard B, Dina D: Moloney murine sarcoma proviral DNA is a transcriptional unit. Nature 288: 665–669 (1980).

    PubMed  Google Scholar 

  2. Bouman H, Mennes AM, Libbenga KR: Transcription in nuclei isolated from tobacco tissues. FEBS Lett 101: 369–372 (1979).

    Article  PubMed  Google Scholar 

  3. Burgess RR: RNA polymerase. Annu Rev Biochem 40: 711–740 (1971).

    Article  PubMed  Google Scholar 

  4. Furter R, Hall BD: Specific transcription and reinitiation of class III genes in wheat embryo nuclei and chromatin. Plant Mol Biol 12: 567–577 (1989).

    Google Scholar 

  5. Hagen G, Guilfoyle TJ: Rapid induction of selective transcription by auxins. Mol Cell Biol 5: 1197–1203 (1985).

    PubMed  Google Scholar 

  6. Hentschel CC, Tata JR: Differential activation of free and engaged RNA polymerase I and II during the resumption of development of dormantArtemia gastrulae. Devel Biol 57: 293–304 (1977).

    Google Scholar 

  7. Hentschel CC, Tata JR: Template-engaged and free RNA polymerases duringXenopus erythroid cell maturation. Devel Biol 65: 496–507 (1978).

    Google Scholar 

  8. Key JL: Modulation of gene expression by auxin. BioEssays 11: 52–58 (1989).

    PubMed  Google Scholar 

  9. Kikuchi M, Imaseki H, Sakai S: Modulation of gene expression in isolated nuclei by auxin-binding proteins. Plant Cell Physiol 30: 765–773 (1989).

    Google Scholar 

  10. Kuhlemeier C, Green PJ, Chua N-H: Regulation of gene expression in higher plants. Annu Rev Plant Physiol 38: 221–257 (1987).

    Google Scholar 

  11. Linsmaier EM, Skoog F: Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18: 100–127 (1965).

    Google Scholar 

  12. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  13. Matrisian LM, Glaichenhaus N, Gesnel M-C, Breathnach R: Epidermal growth factor and oncogenes induce transcription of the same cellular mRNA in rat fibroblasts. EMBO J 4: 1435–1440 (1985).

    PubMed  Google Scholar 

  14. Mennes AM, Bouman H, Van derBurg MPM, Libbenga KR: RNA synthesis in isolated tobacco callus nuclei, and the influence of phytohormones. Plant Sci Lett 13: 329–339 (1978).

    Article  Google Scholar 

  15. Reeve AE, Smith MM, Pigiet V, Huang RCC: Incorporation of purine nucleoside 5′-[γ-S] triphosphates as affinity probes for initiation of RNA synthesisin vitro. Biochemistry 16: 4464–4469 (1977).

    PubMed  Google Scholar 

  16. Reeve AE, Shatkin AJ, Huang RCC: Guanosine 5′-O-(3-thiotriphosphate) inhibits capping of reovirus mRNA. J Biol Chem 257: 7018–7022 (1982).

    PubMed  Google Scholar 

  17. Smith MM, Reeve AE, Huang RCC: Transcription of bacteriophage λ DNAin vitro using purine nucleoside 5′-(γ-S) triphosphates as affinity probes for RNA chain initiation. Biochemistry 17: 493–500 (1978).

    PubMed  Google Scholar 

  18. Tata JR, Baker BS: Specific switching on of silent egg protein genesin vitro by an S-100 fraction in isolated nuclei from maleXenopus. EMBO J 4: 3253–3258 (1985).

    PubMed  Google Scholar 

  19. Thorburn A, Knowland J: A novel nuclear transcription system which responds correctly to cloned estrogen receptor. Nucl Acids Res 16: 10469–10476 (1988).

    PubMed  Google Scholar 

  20. van derLinde PCG, Bouman H, Mennes AM, Libbengta KR: A soluble auxin-binding protein from cultured tobacco tissues stimulates RNA synthesisin vitro. Planta 160: 102–108 (1984).

    Google Scholar 

  21. van derZaal EJ, Mennes AM, Libbenga KR: Auxin-induced rapid changes in translatable mRNAs in tobacco cell suspension. Planta 172: 514–519 (1987).

    Google Scholar 

  22. van derZaal EJ, Memelink J, Mennes AM, Quint A, Libbenga KR: Auxin-induced mRNA species in tobacco cell cultures. Plant Mol Biol 10: 145–157 (1987).

    Google Scholar 

  23. Zhang-Keck Z, Stallcup MR: Optimized reaction conditions and specific inhibitors for initiation of transcription by RNA polymerase II in nuclei from cultured mammalian cells. J Biol Chem 263: 3513–3520 (1988).

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Molecular Biology, Botanical Laboratory, Leiden University, Nonnensteeg 3, 2311 VJ, Leiden, Netherlands

    A. M. Mennes, A. Quint, J. H. Gribnau, C. J. M. Boot, E. J. van der Zaal, A. C. Maan & K. R. Libbenga

Authors
  1. A. M. Mennes
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Quint
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. H. Gribnau
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C. J. M. Boot
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E. J. van der Zaal
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. C. Maan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. K. R. Libbenga
    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

Mennes, A.M., Quint, A., Gribnau, J.H. et al. Specific transcription and reinitiation of 2,4-D-induced genes in tobacco nuclei. Plant Mol Biol 18, 109–117 (1992). https://doi.org/10.1007/BF00018461

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00018461

Key words

Search

Navigation