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.

  • Research Paper
  • Published:

A Genetic System for Rapidly Assessing Herbicides That Compete for the Quinone Binding Site of Photosynthetic Reaction Centers

Bio/Technology volume 7, pages 69–74 (1989)Cite this article

Abstract

To facilitate genetic analysis and manipulation of the bacterial photosynthetic apparatus, a new plasmid vector bearing the reaction center and light harvesting structural genes (puf operon) of Rhodobacter capsulatus was constructed. This plasmid (pU2922) can be conjugated from E. coli strains (tra+) to Rb. capsulatus with high frequency and shows improved fidelity and stability over Co1E1 derivatives. The puf genes may be expressed in trans from pU2922 in Rb. capsulatus deletion backgrounds to assay the effects of directed mutations or to isolate plasmid–borne spontaneous mutations. We have used this genetic system to characterize a total of 27 mutations at five sequence positions in the secondary quinone–binding domain of the reaction center, where six different herbicides assayed in this study act as competitive inhibitors. The broad spectrum of herbicide response ranging from increased resistance to increased sensitivity that is observed in these engineered bacterial strains may provide a sensitive indicator of the interactions of a given herbicide with the binding site. This collection should prove useful for screening new derivatives for their herbicidal activity and for understanding the chemistry of herbicide binding.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Wraight, C.A. 1981. Oxidation-reduction physical chemistry of the acceptor quinone complex in bacterial photosynthetic reaction cen ters: evidence for a new model of herbicide activity. Israel T. Chem. 21:348–354.

    Article  CAS  Google Scholar 

  2. Kyle, D.J. 1985. The 32,000 dalton QB protein of photosystem II. Photochem. Photobiol. 41:107–116.

    Article  CAS  Google Scholar 

  3. Erickson, J.M., Rahire, M., Metz, L., and Rochaix, J.-D. 1985. Herbicide resistance and cross-resistance: changes at three distinct sites in the herbicide-binding protein. Science 228:204–207.

    Article  CAS  Google Scholar 

  4. Johanningmeier, U., Bodner, U. and Wildner, G.F. 1987. A new mutation in the gene encoding for the herbicide-binding protein in Chlamydomonas. FEBS Lett. 211:221–224.

    Article  CAS  Google Scholar 

  5. Rochaix, J.-D. and Erickson, J. 1988. Function and assembly of photosystem II: genetic and molecular analysis. Trends Biochem. Sci. 13:56–59.

    Article  CAS  Google Scholar 

  6. Sinning, I. and Michel, H. 1987. Sequence analysis of mutants from Rhodopseudomonas viridis resistant to the herbicide terbutryn. Z. Natur-forsch. 42c:751–754.

    Google Scholar 

  7. Gilbert, C.W., Williams, J.G.K., Williams, K.A.L., and Arntzen, C.J. 1985. Herbicide action in photosynthetic bacteria, p. 67–71. In: Molecular Biology of the Photosynthetic Apparatus. K. E. Steinback, S. Bonitz, C. J. Arntzen, and L. Bogorad (Eds.). Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  8. Paddock, M.L., Williams, J.C., Rongey, S.H., Abresch, E.C., Feher, G. and Okamura, M.Y. 1987. Characterization of three herbicide resistant mutants of Rhodopseudomonas sphaeroides 2.4.1: structure-function relationship. Vol III, p. 775–778. In: Progress in Photosynthetic Research, J. Biggins (Ed.) Martinus Nijhoff, Dordrecht.

    Chapter  Google Scholar 

  9. Bylina, E.J. and Youvan, D.C. 1987. Genetic engineering of herbicide resistance: saturation mutagenesis of isoleucine 229 of the reaction center L subunit. Z. Naturforsch. 42c:769–774.

    Article  Google Scholar 

  10. Michel, H., Epp, O., and Deisenhofer, J. 1986. Pigment-protein interactions in the photosynthetic reaction centre from Rhodopseudomonas viridis. EMBO J. 5:2445–2451.

    Article  CAS  Google Scholar 

  11. Bylina, E.J. and Youvan, D.C. 1988. Directed mutations affecting spectroscopic and electron transfer properties of the primary donor in the photosynthetic reaction center. Proc. Natl. Acad. U.S.A. In press.

  12. Scolnik, P.A. and Marrs, B.L. 1987. Genetic research with photosynthetic bacteria. Ann. Rev. Microbiol. 41:703–726.

    Article  CAS  Google Scholar 

  13. Youvan, D.C., Ismail, S. and Bylina, E.J. 1985. Chromosomal deletion and plasmid complementation of the photosynthetic reaction center and light harvesting genes from Rhodopseudomonas capsulata. Gene 38:19–30.

    Article  CAS  Google Scholar 

  14. Bylina, E.J., Ismail, S. and Youvan, D.C. 1986. Plasmid pU29, a vehicle for mutagenesis of the photosynthetic puf operon in Rhodopseudomonas capsulata. Plasmid 16:175–181.

    Article  CAS  Google Scholar 

  15. Taylor, D.P., Cohen, S.N., Clark, W.G. and Marrs, B.L. Alignment of genetic and restriction maps of the photosynthesis region of the Rhodopseudomonas capsulata chromosome by a conjugation-mediated marker rescue technique. J. Bacteriol. 154:580–590.

  16. Bylina, E.J. 1988. Ph.D. Dissertation. State University of New York at Stony Brook.

  17. Ditta, G., Stanfield, S., Corbin, D., and Helinski, D.R. 1980. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc. Natl. Acad. U.S.A. 77:7347–7351.

    Article  CAS  Google Scholar 

  18. Simon, R., Priefer, U., and Pühler, A. 1983. A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Bio/Technology 1:784–791.

    Article  CAS  Google Scholar 

  19. Youvan, D.C., Bylina, E.J., Alberti, M., Begusch, H., and Hearst, J.E. 1984. Nucleotide and deduced polypeptide sequences of the photosynthetic reaction-center, B870 antenna, and flanking polypeptides from R. capsulata. Cell 37:949–957.

    Article  CAS  Google Scholar 

  20. Prince, R.C. and Youvan, D.C. 1987. Isolation and spectroscopic properties of photochemical reaction centers from Rhodobacter capsu-latus. Biochim. Biophys. Acta 890:286–291.

    Article  CAS  Google Scholar 

  21. Allen, J.P., Feher, G., Yeates, T.O., Rees, D.C., Deisenhofer, J., Michel, H., and Huber, R. 1986. Structural homology of reaction centers from Rhodopseudomonas sphaeroides and Rhodopseudomonas viridis as determined by x-ray diffraction. Proc. Natl. Acad. Sci. U.S.A. 83:8589–8593.

    Article  CAS  Google Scholar 

  22. Chang, C.-H., Tiede, D., Tang, J., Smith, U., Norris, J., and Schiffer, M. 1986. Structure of Rhodopseudomonas sphaeroides R-26 reaction center. FEBS Lett. 205:82–86.

    Article  CAS  Google Scholar 

  23. Yadav, N., McDevitt, R.E., Benard, S., and Falco, S.C. 1986. Single amino acid substitutions in the enzyme acetolactate synthase confer resistance to the herbicide sulfometuron methyl. Proc. Natl. Acad. Sci. U.S.A. 83:4418–4422.

    Article  CAS  Google Scholar 

  24. Sinning, I. and Michel, H. 1987. Terbutryn-resistant mutants of Rhodopseudomonas viridis are double mutated. Vol III, p. 771–773. In: Progress in Photosynthetic Research, J. Biggins (Ed.). Martinus Nijhoff, Dordrecht.

    Chapter  Google Scholar 

  25. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  26. Zoller, M.J. and Smith, M. 1984. Oligonucleotide-directed mutagenesis: a simple method using two oligonucleotide primers and a single-stranded DNA template. DNA 3:479–488.

    Article  CAS  Google Scholar 

  27. Youvan, D.C., Elder, J.T., Sandlin, D.E., Zsebo, K., Alder, D.P., Panopoulos, N.J., Marrs, B.L., and Hearst, J.E. 1982. R-prime site-directed transposon Tn7 mutagenesis of the photosynthetic apparatus in Rhodopseudomonas capsulata. J. Mol. Biol. 162:19–41.

    Article  Google Scholar 

  28. Yen, H.-C. and Marrs, B.L. 1977. Growth of Rhodopseudomonas capsulata under aerobic dark conditions with dimethyl sulfoxide. Arch. Biochem. Biophys. 181:411–418.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Douglas C. Youvan: Corresponding Author.

Authors and Affiliations

  1. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, 02139

    Edward J. Bylina, Raffael V. M. Jovine & Douglas C. Youvan

Authors
  1. Edward J. Bylina
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Raffael V. M. Jovine
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Douglas C. Youvan
    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

Bylina, E., Jovine, R. & Youvan, D. A Genetic System for Rapidly Assessing Herbicides That Compete for the Quinone Binding Site of Photosynthetic Reaction Centers. Nat Biotechnol 7, 69–74 (1989). https://doi.org/10.1038/nbt0189-69

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0189-69

Search

Advanced 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