Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

A light-switchable gene promoter system

Abstract

Regulatable transgene systems providing easily controlled, conditional induction or repression of expression are indispensable tools in biomedical and agricultural research and biotechnology. Several such systems have been developed for eukaryotes1,2,3,4,5,6. Most of these rely on the administration of either exogenous chemicals or heat shock. Despite the general success of many of these systems, the potential for problems, such as toxic, unintended, or pleiotropic effects of the inducing chemical or treatment, can impose limitations on their use. We have developed a promoter system that can be induced, rapidly and reversibly, by short pulses of light. This system is based on the known red light–induced binding of the plant photoreceptor phytochrome to the protein PIF3 and the reversal of this binding by far-red light7,8. We show here that yeast cells expressing two chimeric proteins, a phytochrome–GAL4-DNA-binding-domain fusion and a PIF3–GAL4-activation-domain fusion, are induced by red light to express selectable or “scorable” marker genes containing promoters with a GAL4 DNA-binding site, and that this induction is rapidly abrogated by subsequent far-red light. We further show that the extent of induction can be controlled precisely by titration of the number of photons delivered to the cells by the light pulse. Thus, this system has the potential to provide rapid, noninvasive, switchable control of the expression of a desired gene to a preselected level in any suitable cell by simple exposure to a light signal.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Light-responsive gene promoter system.
Figure 2: Photoreversible activation of reporter gene expression.

References

  1. Blau, H.M. & Rossi, F.M.V. Tet B or not tet B: advances in tetracycline-inducible gene expression. Proc. Natl. Acad. Sci. USA 96, 797–799 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Gatz, C. Chemically inducible promoters in transgenic plants. Curr. Opin. Biotech. 7, 168–172 (1996).

    Article  CAS  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  4. Lewandoski, M. Conditional control of gene expression in the mouse. Nature Rev. Genet. 2, 743–755 (2001).

    Article  CAS  PubMed  Google Scholar 

  5. Mills, A.A. Changing colors in mice: an inducible system that delivers. Genes Dev. 15, 1461–1467 (2001).

    Article  CAS  PubMed  Google Scholar 

  6. Roslan, H.A. et al. Characterization of the ethanol-inducible alc gene-expression system in Arabidopsis thaliana. Plant J. 28, 225–235 (2001).

    Article  CAS  PubMed  Google Scholar 

  7. Ni, M., Tepperman, J.M. & Quail, P.H. Binding of phytochrome B to its nuclear signaling partner PIF3 is reversibly induced by light. Nature 400, 781–784 (1999).

    Article  CAS  PubMed  Google Scholar 

  8. Zhu, Y., Tepperman, J.M., Fairchild, C.D. & Quail, P.H. Phytochrome B binds with greater apparent affinity than phytochrome A to the basic helix-loop-helix factor PIF3 in a reaction requiring the PAS domain of PIF3. Proc. Natl. Acad. Sci. USA 97, 13419–13424 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Quail, P.H. Phytochrome photosensory signalling networks. Nature Rev. Mol. Cell Biol. 3, 85–93 (2002).

    Article  CAS  Google Scholar 

  10. Quail, P.H. An emerging molecular map of the phytochromes. Plant Cell Environ. 20, 657–665 (1997).

    Article  CAS  Google Scholar 

  11. Phizicky, E.M. & Fields, S. Protein-protein interactions: methods for detection and analysis. Microbiol. Rev. 59, 94–123 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Gambetta, G.A. & Lagarias, J.C. Genetic engineering of phytochrome biosynthesis in bacteria. Proc. Natl. Acad. Sci. USA 98, 10566–10571 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Smith, H. & Jackson, G.M. Rapid phytochrome regulation of wheat seedling extension. Plant Physiol. 84, 1059–1062 (1987).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Ronicke, V., Graulich, W., Mumberg, D., Muller, R. & Funk, M. Use of conditional promoters for expression of heterologous proteins in Saccharomyces cerevisiae. Meth. Enzymol. 283, 313–322 (1997).

    Article  CAS  Google Scholar 

  15. Parks, B.M. & Quail, P.H. Phytochrome-deficient hy1 and hy2 long hypocotyl mutants of Arabidopsis are defective in phytochrome chromophore biosynthesis. Plant Cell 3, 1177–1186 (1991).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Boylan, M.T. & Quail, P.H. Phytochrome A overexpression inhibits hypocotyl elongation in transgenic Arabidopsis. Proc. Natl. Acad. Sci. USA 88, 10806–10810 (1991).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Harper, J.W., Adami, G.R., Wei, N., Keyomarsi, K. & Elledge, S.J. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell 75, 805–816 (1993).

    Article  CAS  PubMed  Google Scholar 

  18. Scheer, H. Model compounds for the phytochrome chromophore. in Techniques in photomorphogenesis. (eds Smith, H. & Holmes, M.G.) 227–256 (Academic Press, New York, 1984).

    Google Scholar 

  19. James, P., Halliaday, J. & Craig, E.A. Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics 144, 1425–1436 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Parks, B.M. & Quail, P.H. hy8, a new class of Arabidopsis long hypocotyl mutants deficient in functional phytochrome A. Plant Cell 5, 39–48 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank the members of our laboratory for support and discussions. This work was supported by grants from the Department of Energy Basic Energy Sciences (number DE-FG03-87ER13742), the National Institutes of Health (number GM47475), and the US Department of Agriculture Current Research Information Service (number 5335-21000-010-00D).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Peter H. Quail.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Shimizu-Sato, S., Huq, E., Tepperman, J. et al. A light-switchable gene promoter system. Nat Biotechnol 20, 1041–1044 (2002). https://doi.org/10.1038/nbt734

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt734

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing