Skip to main content
SpringerLink
Log in

Absence of PsaC subunit allows assembly of photosystem I core but prevents the binding of PsaD and PsaE in Synechocystis sp. PCC6803

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

In photosystem I (PSI) of oxygenic photosynthetic organisms the psaC polypeptide, encoded by the psaC gene, provides the ligands for two [4Fe-4S] clusters, FA and FB. Unlike other cyanobacteria, two different psaC genes have been reported in the cyanobacterium Synechocystis 6803, one (copy 1) with a deduced amino acid sequence identical to that of tobacco and another (copy 2) with a deduced amino acid sequence similar to those reported for other cyanobacteria. Insertion of a gene encoding kanamycin resistance into copy 2 resulted in a photosynthesis-deficient strain, CDK25, lacking the PsaC, PsaD and PsaE polypeptides in isolated thylakoid membranes, while the PsaA/PsaB and PsaF subunits were found. Growth of the mutant cells was indistinguishable from that of wild-type cells under light-activated heterotrophic growth (LAHG). A reversible P700+ signal was detected by EPR spectroscopy in the isolated thylakoids during illumination at low temperature. Under these conditions, the EPR signals attributed to FA and FB were absent in the mutant strain, but a reversible Fx signal was present with broad resonances at g=2.079, 1.903, and 1.784. Addition of PsaC and PsaD proteins to the thylakoids gave rise to resonances at g=2.046, 1.936, 1.922, and 1.880; these values are characteristic of an interaction-type spectrum of FA - and FB -. In room-temperature optical spectroscopic analysis, addition of PsaC and PsaD to the thylakoids also restored a 30 ms kinetic transient which is characteristic of the P700+ [FA/FB]- backreaction. Expression of copy 1 was not detected in cells grown under LAHG and under mixotrophic conditions. These results demonstrate that copy 2 encodes the PsaC polypeptide in PSI in Synechocystis 6803, while copy 1 is not involved in PSI; that the PsaC polypeptide is necessary for stable assembly of PsaD and PsaE into PSI complex in vivo; and that PsaC, PsaD and PsaE are not needed for assembly of PsaA-PsaB dimer and electron transport from P700 to Fx.

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. Anderson SL, McIntosh L: Light-activated heterotrophic growth of the cyanobacterium Synechocystic sp. strain PCC 6803: a blue-light-requiring process. J Bact 173: 2761–2767 (1991).

    Google Scholar 

  2. Anderson SL, McIntosh L: Partial conservation of the 5′ ndhE-psaC-ndhD 3′ gene arrangement of chloroplasts in the cyanobacterium Synechocystis sp. PCC 6803: implications for NDH-D function in cyanobacteria and chloroplasts. Plant Mol Biol 16: 497–499 (1991).

    Google Scholar 

  3. Chitnis VP, Xu Q, Yu L, Golbeck JH, Nakamoto H, Xie DL, Chitnis PR: Targeted inactivation of the gene psaL encoding a subunit of photosystem I of the cyanobacterium Synechocystis sp PCC 6803. J Biol Chem 268: 11678–11684 (1993).

    Google Scholar 

  4. Feinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).

    Google Scholar 

  5. Fish LE, Kück U, Bogorad L: Two partially homologous adjacent light-inducible maize chloroplast genes encoding polypeptides of the P700 chlorophyll a-protein complex of photosystem I. J Biol Chem 260: 1413–1421 (1985).

    Google Scholar 

  6. Golbeck JH: Structure and function of photosystem I. Annu Rev Plant Physiol Plant Mol Biol 43: 293–324 (1992).

    Google Scholar 

  7. Golbeck JH: Photosystem I in Cyanobacteria. In: Bryant DA (ed) The Molecular Biology of Cyanobacteria, pp. 179–220. Kluwer Academic Publishers. Dordrecht, Netherlands (1994).

    Google Scholar 

  8. Henry RL, Li M, Guikema JA: A comparison of the PSI polypeptide organization on soybean and the cyanobacterium, Anacystis nidulans. In: Baltscheffsky M (ed) Current Research in Photosynthesis. vol. 2, pp. 567–570. Kluwer Academic Publishers. Dordrecht, Netherlands (1990).

    Google Scholar 

  9. HøJ PB, Svendsen I, Scheller HV, Møller BR: Identification of a chloroplast-encoded 9-kDa polypeptide as a 2[4Fe-4S] protein carrying centers A and B of photosystem I. J Biol Chem 262: 12676–12684 (1987).

    Google Scholar 

  10. Jansson C, Debus RJ, Osiewacz HD, Gurevitz M, McIntosh L: Construction of an obligate photoheterotrophic mutant of the cyanobacterium Synechocystis 6803. Plant Physiol 85: 1021–1025 (1987).

    Google Scholar 

  11. Li N, Warren PV, Golbeck JH, Frank G, Zuber H, Bryant DA: Polypeptide composition of the photosystem I complex and the photosystem I core protein from Synechococcus sp. PCC 6301. Biochim Biophys Acta 1059: 215–225 (1991).

    Google Scholar 

  12. Li N, Zhao J, Warren PV, Warden JT, Bryant DA, Golbeck JH: PsaD is required for the stable binding of PsaC to the photosystem I core protein of Synechococcus sp. PCC 6301. Biochemistry 30: 7863–7872 (1991).

    Google Scholar 

  13. Lichtenthaler HK: Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Meth Enzymol 148: 350–382 (1987).

    Google Scholar 

  14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275 (1951).

    Google Scholar 

  15. Mannan RM, Pakrasi HB, Sonoike K: The PsaC protein is necessary for the stable association of the PsaD, PsaE, and PsaL proteins in the photosystem I complex: analysis of a cyanobacterial mutant strain. Arch Biochem Biophys 315: 68–73 (1994).

    Google Scholar 

  16. Mannan RM, Whitmarsh J, Nyman P, Pakrasi HB: Directed mutagenesis of an iron-sulfur protein of the photosystem I complex in the filamentous cyanobacterium Anabaena variabilis ATCC 29413. Proc Natl Acad Sci USA 88: 10168–10172 (1991).

    Google Scholar 

  17. Noren GH, Boerner RJ, Barry BA: EPR characterization of an oxygen-evolving photosystem II preparation from the transformable cyanobacterium Synechocystis 6803. Biochemistry 30: 3943–3950 (1991).

    Google Scholar 

  18. Oh-oka H, Takahashi Y, Kuriyama K, Saeki K, Matsubara H: The protein responsible for center A/B in spinach photosystem I: isolation with iron-sulfur cluster(s) and complete sequence analysis. J Biochem 103: 962–968 (1988).

    Google Scholar 

  19. Oka A, Sugisaki H, Takanami M: Nucleotide sequence of the kanamycin resistance transposon Tn903. J Mol Biol 174: 217–226 (1981).

    Google Scholar 

  20. Parrett KG, Mehari T, Warren PV, Golbeck JH: Purification and properties of the intact P700 and Fx-containing photosystem I core protein. Biochim Biophys Acta 973: 324–332 (1989).

    Google Scholar 

  21. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  22. Shen G, Boussiba S, Vermaas WFJ: Synechocystis sp PCC 6803 strains lacking photosystem I and phycobilisome function. Plant Cell 5: 1853–1863 (1993).

    Google Scholar 

  23. Smart LB, Anderson SL, McIntosh L: Targeted genetic inactivation of the photosystem I reaction center in the cyanobacterium Synechocystis sp PCC 6803. EMBO J 10: 3289–3296 (1991).

    Google Scholar 

  24. Smart LB, Warren PV, Golbeck JH, McIntosh L: Mutational analysis of the structure and biogenesis of the photosystem I reaction center in the cyanobacterium Synechocystis sp PCC 6803. Proc Natl Acad Sci USA 90: 1132–1136 (1993).

    Google Scholar 

  25. Steinmüller K: Identification of a second psaC gene in the cyanobacterium Synechocystis sp PCC 6803. Plant Mol Biol 20: 997–1001 (1992).

    Google Scholar 

  26. takahashi Y, Goldschmidt-Clermont M, Soen SY, Franzen, Rochaix JD: Directed chloroplast transformation in Chlamydomonas reinhardtii: insertional inactivation of the psaC gene encoding the iron sulfur protein destabilizes photosystem I. EMBO J 10: 2033–2040 (1991).

    Google Scholar 

  27. Vermaas WFJ, Ikeuchi M, Inoue Y: Protein composition of the photosystem II core complex in genetically engineered mutants of the cyanobacterium Synechocystis sp PCC 6803. Photosynth Res 17: 97–113 (1988).

    Google Scholar 

  28. Vieira J, Messing J: The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19: 259–268 (1982).

    Google Scholar 

  29. Warren PV, Smart LB, McIntosh L, Golbeck JH: Sitedirected conversion of cysteine 565 to serine in PsaB of photosystem I results in the assembly of [3Fe-4S] and [4Fe-4S] clusters in Fx. A mixed-ligand [4Fe-4S] cluster is capable of electron transfer to FA and FB. Biochemistry 32: 4411–4419 (1993).

    Google Scholar 

  30. Williams JGK: Construction of specific mutations in photosystem II photosynthetic reaction center by genetic engineering methods in Synechocystis 6803. Meth Enzymol 167: 766–778 (1988).

    Google Scholar 

  31. Wynn RM, Malkin R: Interaction of plastocyanin with photosystem I: a chemical cross-linking study of the polypeptide that binds plastocyanin. Biochemistry 27: 5863–5869 (1988).

    Google Scholar 

  32. Yu L, Zhao J, Mühlenhoff, Bryant DA, Golbeck JH: PsaE is required for in vivo cyclic electron flow around photosystem I in the cyanobacterium Synechococcus sp. PCC 7002. Plant Physiol 103: 171–180 (1993).

    Google Scholar 

  33. Zilber AL, Malkin R: Ferredoxin cross-links to a 22 kD subunit of photosystem I. Plant Physiol 88: 810–814 (1988).

    Google Scholar 

Download references

Author information

Author notes

  1. Lawrence B. Smart

    Present address: Department of Vegetable Crops, Mann Laboratory, University of California, 95616, Davis, CA, USA

Authors and Affiliations

  1. DOE Plant Research Laboratory, Michigan State University, 48824, East Lansing, MI, USA

    Jianping Yu, Lawrence B. Smart & Lee McIntosh

  2. Department of Biochemistry, University of Nebraska, 68583, Lincoln, NE, USA

    Yean-Sung Jung & John Golbeck

Authors
  1. Jianping Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lawrence B. Smart
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yean-Sung Jung
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. John Golbeck
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lee McIntosh
    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

Yu, J., Smart, L.B., Jung, YS. et al. Absence of PsaC subunit allows assembly of photosystem I core but prevents the binding of PsaD and PsaE in Synechocystis sp. PCC6803. Plant Mol Biol 29, 331–342 (1995). https://doi.org/10.1007/BF00043656

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00043656

Key words

Search

Navigation