Skip to main content
SpringerLink
Log in

Characterization of photosystem II mutants of Chlamydomonas reinhardii lacking the psbA gene

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Summary

We have examined 78 chloroplast mutants of Chlamydomonas reinhardii lacking photosystem II activity. Most of them are unable to synthesize the 32 Kdalton protein. Analysis of 22 of these mutants reveals that they have deleted both copies of the psbA gene (which codes for the 32 Kdalton protein) in their chloroplast genome. Although these mutants are able to synthesize and to integrate the other photosystem II polypeptides in the thylakoid membranes, they are unable to assemble a stable functional photosystem II complex. The 32 Kprotein appears therefore to play an important role not only in photosystem II function, but also in stabilizing this complex.

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

  1. Arntzen CJ, Ditto CL, Brewer PE: Chloroplast membrane alterations in traizine-resistant Amaranthus retroflexus biotypes. Proc Natl Acad Sci USA 76:278–282, 1979.

    Google Scholar 

  2. Arntzen CJ, Pfister K, Steinback KE: The mechanism of chloroplast triazine resistance: alteration in the site of herbicide action. In: Le Baron HM, Gressel J (eds) Herbicide resistance in plants. Wiley-Interscience Publications, New York, 1982, pp 184–214.

    Google Scholar 

  3. Bennoun P, Masson A, Piccioni R, Chua NH: Uniparental mutants of Chlamydomonas reinhardii defective in photosynthesis. In: Akoyunoglou A, Argyroudi-Akoyunoglou JH (eds)_ Chloroplast Development. Elsevier/North-Holland Biomedical Press, 1978, pp 721–726.

  4. Bennoun P, Masson A, Delosme M: A method for complementation analysis of nuclear and chloroplast mutants of photosynthesis in Chlamydomonas. Genetics 95:39–47, 1980.

    Google Scholar 

  5. Bennoun P, Diner B, Wollman F, Schmidt G, Chua NH: Thylakoid polypeptides associated with the photosystem II in Chlamydomonas reinhardii: comparisons of photosystem II mutants and particles. In: Akoyunoglou G (eds) Structure and Molecular Organization of the photosynthetic apparatus. Balaban international Series, 1981, pp 839–849.

  6. Bennoun P, Delepelaire P: Isolation of photosynthesis mutants in Chlamydomonas. In: Edelman M, Hallick RB, Chua NH (eds) Methods in Chloroplast Molecular Biology. Elsevier Biomedical Press, 1982, pp 25–38.

  7. Chua NH, Bennoun P: Thylakoid membrane polypeptides of Chlamydomonas reinhardii: Wild type and mutant strains deficient in photosystem II reaction center. Proc Natl Acad Sci USA 72:2175–2179 1975.

    Google Scholar 

  8. Covey C, Richardson D, Carbon J: A method for the detection of restriction sites in bacterial plasmid deoxyribonucleic acid. Molec Gen Genet 145:155–158, 1976.

    Google Scholar 

  9. Davis RW, Thomas M, Cameron J, St John TP, Scherrer S, Padgett RA: Rapid DNA isolation for enzymatic and hybridization analysis. In Meth in Enzymol 65:404–411, 1980.

    Google Scholar 

  10. Delepelaire P: Partial characterization of the biosynthesis and integration of the photosystem II reaction centers in the thylakoid membrane of Chlamydomonas reinhardii. EMBO J 3:701–706, 1984.

    Google Scholar 

  11. Delepelaire P, Chua NH: Lithium dodecyl sulfate/polyacrylamide gel electrophoresis of thylakoid membranes at 4°C; Characterizations of two additional chlorophyll a-protein complexes. Proc Natl Acad Sci USA 76:111–115, 1979.

    Google Scholar 

  12. Delepelaire P, Wollman F: Correlation between fluorescent and phosphorylation changes in vivo: Kinetic analysis. Biochem Biophys Acta, in press, 1985.

  13. Erickson JM, Rahire M, Rochaix J-D: C. reinhardii gene fort the Mr 32000 protein of photosystem II contains four large introns and is located entirely within the chloroplast inverted repeat. EMBO J 3:2753–2762.

  14. Erickson JM, Rahire M, Bennoun P, Delepelaire P, Diner B, Rochaix J-D: Herbicide resistance in C. reinhardii results from a mutation in the chloroplast gene for the 32 kilodalton protein of photosystem II. Proc Natl Acad Sci 81:3617–3621, 1984.

    Google Scholar 

  15. Erickson JM, Rahire M, Rochaix J-D, Mets L: Herbicide resistance and crossresistance: Changes at three distinct sites in the herbicide-binding protein. Science 228:204–207, 1985.

    Google Scholar 

  16. Erickson JM, Delepelaire P, Rochaix J-D: Analysis of genes encoding two photosystem II proteins of the 30–34 KD size class. In: Arntzen CJ, Bonitz S, Bogorad L, Steinback K (eds) Molecular Biology of the Photosynthetic Apparatus. Cold Spring Harbour, NY, 1985, in press.

  17. Galloway R, Mets L: Atrazine, bromacil and diuron resistance in Chlamydomonas: A single non-mendelian genetic locus controls the structure of the thylakoid binding site. Plant Physiol 74:469–474, 1984.

    Google Scholar 

  18. Girard J, Chua NH, Bennoun P, Schmidt G, Delosme M: Studies on mutants deficient in photosystem I reaction centers in Chlamydomonas reinhardii. Current Genet 2:215–221, 1980.

    Google Scholar 

  19. Glisin V, Crkvenjakov R, Byns C: Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry 13:2633–2637, 1974.

    Google Scholar 

  20. Goloubinoff P, Edelman M, Hallick RB: Chloroplastcoded atrazine resistance in Solanum nigrum: psbA loci from susceptible and resistant biotypes are isogenic except for a single codon change. Nucl Acids Res 12:5489–9496.

  21. Gorman DS, Levine RP: Cytochrome F and plastocyanin: Their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardii. Proc Natl Acad Sci USA 54:1665–1669, 1965.

    Google Scholar 

  22. Hirschberg J, McIntosh L: Molecular basis of herbicide resistance in Amaranthus hybridus. Science 222:1346–1348, 1983.

    Google Scholar 

  23. Karabin GD, Farley M, Hallick RB: Chloroplast gene for Mr 32000 polypeptide of photosystem II in Euglena gracilis is interrupted by four introns with conserved boundary sequences. Nucleic Acids Res 12:5801–5812, 1984.

    Google Scholar 

  24. Keller M, Stutz E: Structure of the Euglena gracilis chloroplast gene (psbA) coding for the 32 KD protein of photosystem II. FEBS Letts 175:173–177, 1984.

    Google Scholar 

  25. Kyle DJ, Ohad I, Arntzen CJ: Membrane protein damage and repair: Selective loss of a quinone-protein function in chloroplast membranes. Proc Natl Acad Sci USA 81:4070–4074, 1984.

    Google Scholar 

  26. Mattoo AK, Hoffman-Falk H, Marder JB, Edelman M: Regulation of protein metabolism: coupling of photosynthetic electron transport to in vivo degradation of the rapidly metabolized 32 kDa protein of the chloroplast membranes. Proc Natl Acad Sci USA 81:1380–1384, 1984.

    Google Scholar 

  27. Palmer JD, Boynton J, Gilham NW, Harris EH: Evolution and recombination of the large inverted repeat in Chlamydomonas chloroplast DNA. In: Arntzen C, Bogorad L, Bonitz S, Steinback K (eds) Cold Spring Harbor Symp. on Molecular Biology of the Photosynthetic apparatus. 1985, in press.

  28. Pfister K, Steinback KE, Gardner G, Arntzen CJ: Photoaffinity labeling of an herbicide receptor protein in chloroplast membranes. Proc Natl Acad Sci USA 78:981–985, 1981.

    Google Scholar 

  29. Piccioni P, Bennoun P, Chua NH: A nuclear mutant of Chlamydomonas reinhardii deficient in photosynthetic phosphorylation: characterization of the algal coupling factor ATPase. Eur J Biochem 117:93–102, 1981.

    Google Scholar 

  30. Rave N, Crkvenjakov R, Boedtker H: Identification of procollagen mRNAs transferred to diazobenzyloxymethyl paper from formaldehyde agarose gels. Nucleic Acids Res 6:3559–3567, 1979.

    Google Scholar 

  31. Rochaix JD: Restriction endonuclease map of the chloroplast DNA of Chlamydomonas reinhardii. J Mol Biol 126:597–617, 1978.

    Google Scholar 

  32. Rochaix JD: Organization, function and expression of the chloroplast DNA of Chlamydomonas reinhardii. Experientia 37: 323–332, 1981.

    Google Scholar 

  33. Rochaix JD, van Dillewijn J: Transformation of the green alga Chlamydomonas reinhardii with yeast DNA. Nature 296:70–72, 1982.

    Google Scholar 

  34. Rochaix JD, Dron M, Rahire M, Malnoe P: Sequence homology between the 32 K dalton and D2 chloroplast membrane polypeptides of Chlamydomonas reinhardii. Plant Molec Biol 3:363–370, 1984.

    Google Scholar 

  35. Southern EM: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517, 1975.

    Google Scholar 

  36. Spreitzer R, Mets L: Photosynthesis-deficient mutants of Chlamydomonas reinhardii with associated light-sensitive phenotypes. Plant Physiol 67:565–569, 1981.

    Google Scholar 

  37. Thomas PS: Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205, 1980.

    Google Scholar 

  38. Williams JC, Steiner LA, Ogden R, Simon M, Feher G: Primary structure of the M subunit of the reaction center from Rhodopseudomonas sphaeroides. Proc Natl Acad Sci USA 80:6505–6509, 1983.

    Google Scholar 

  39. Wollman FA, Delepelaire P: Correlation between changes in light energy distribution and changes in thylakoid membrane polypeptide phosphorylation in Chlamydomonas reinhardii. J Cell Biol 98:1–7, 1984.

    Google Scholar 

  40. Youvan D, Bylina E, Alberti M, Begusch H, Hearst JE: Nucleotide and deduced polypeptide sequences of the photosynthetic reaction center B870 antennae and flanking polypeptides from Rsp. capsulata. Cell 37:949–957, 1984.

    Google Scholar 

  41. Zurawski G, Bohnert HJ, Whitfeld PR, Bottomley W: Nucleotide sequence of the gene for the Mr 32000 thylakoid membrane protein from Spinacia oleracea and Nicotiana debneyi predicts a totally conserved primary translation product of Mr 38950. Proc Natl Acad Sci USA 79:7699–7703, 1982.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut de Biologie Physico-chimique, 75005, Paris, France

    Pierre Bennoun, Jacqueline Girard-Bascou, Yves Pierre & Monique Delosme

  2. Department of Molecular and Plant Biology, University of Geneva, 1211, Geneva 4, Switzerland

    Muriel Spierer-Herz, Jeanne Erickson & J. -D. Rochaix

Authors
  1. Pierre Bennoun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Muriel Spierer-Herz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jeanne Erickson
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jacqueline Girard-Bascou
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yves Pierre
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Monique Delosme
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J. -D. Rochaix
    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

Bennoun, P., Spierer-Herz, M., Erickson, J. et al. Characterization of photosystem II mutants of Chlamydomonas reinhardii lacking the psbA gene. Plant Mol Biol 6, 151–160 (1986). https://doi.org/10.1007/BF00021484

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00021484

Keywords

Search

Navigation