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Site-specific mutations in the D1 polypeptide affect the susceptibility of Synechocystis 6803 cells to photoinhibition

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Abstract

Photoinhibition of photosystem II in the cyanobacterium Synechocystis 6803 was followed after site-specific mutagenesis of the D1 polypeptide. Mutations were created in the stromal/cytosolic loop connecting helices D and E. Two mutations E243K and CA1, a deletion of the three glutamates 242–244 and a substitution Q241H, were made in the putative cleavage area of the D1 polypeptide. A third mutation E229D was made in the PEST-like sequence. Mutants and control cells were illuminated and FV/FM was recorded. Compared to the control, the mutants were less photoinhibited. Fluorescence relaxation after a single flash was delayed in CA1. Restoration of FV/FM after photoinhibition in the mutants was totally dependent on protein synthesis while control cells were able to recover partially also when protein synthesis was inhibited. In addition, the protein synthesis-dependent recovery of CA1 was slowed down. Our results indicate a correlation between the mutated amino acids and photoinhibition of photosystem II.

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References

  1. Aro E-M, Hundal T, Carlberg I, Andersson B: In vitro studies on light-induced inhibition of Photosystem II and D1-protein degradation at low temperatures. Biochim Biophys Acta 1019: 269–275 (1990).

    Google Scholar 

  2. Barber J, Andersson B: Too much of a good thing: light can be bad for photosynthesis. Trends Biochem Sci 17: 61–66 (1992).

    Google Scholar 

  3. Casanova JL, Pannetier C, Jaulin C, Kourilsky P: Optimal conditions for directly sequencing double-stranded PCR products with Sequenase. Nucl Acids Res 18: 4028 (1990).

    Google Scholar 

  4. Davis LG, Dibner MD, Battley JF: Basic methods in molecular biology. New York, NY: Elsevier Science Publishers (1986).

    Google Scholar 

  5. De Las Rivas J, Andersson B, Barber J: Two sites of primary degradation of the D1-protein induced by acceptor or donor side photoinhibition in photosystem II core complexes. FEBS Lett 301: 246–252 (1992).

    Google Scholar 

  6. Erickson JM, Pfister K, Rahire M, Togasaki RK, Mets L, Rochaix J-D: Molecular and biophysical analysis of herbicide-resistant mutants of Chlamydomonas reinhardtii: Structure-function relationship of the photosystem II D1 polypeptide. The Plant Cell 1: 361–371 (1989).

    Google Scholar 

  7. Giersch C, Krause GH: A simple model relating photoinhibitory fluorescence quenching in chloroplasts to a population of altered Photosystem II reaction centers. Photosynth Res 30: 115–121 (1991).

    Google Scholar 

  8. Gong H, Ohad I. The PQ/PQH2 ratio and occupancy of photosystem II-QB site by plastoquinone control the degradation of D1 protein during photoinhibition in vivo. J Biol Chem 266: 21293–21299 (1991).

    Google Scholar 

  9. Govindjee: Photosystem II heterogeneity: the acceptor side. Photosynth Res 25: 151–160 (1990).

    Google Scholar 

  10. Greenberg BM, Gaba V, Mattoo AK, Edelman M: Identification of a primary in vivo degradation product of the rapidly-turning-over 32 kd protein of photosystem II. EMBO J: 2865–2869 (1987).

  11. Haag E, Gleiter HM, Renger G: Effects of photoinhibition on the PS II acceptor side including the endogenous high spin Fe2+ in thylakoids, PS II membrane fragments and PS II core complexes. Photosynth Res 31: 113–126 (1992).

    Google Scholar 

  12. Hundal T, Aro E-M, Carlberg I, Andersson B: Restoration of light-induced photosystem II inhibition without de novo protein synthesis. FEBS Lett 267: 203–206 (1990).

    Google Scholar 

  13. Jansson C, Debus RJ, Osiewacz HD, Gurevitz M, McIntosh L: Construction of an oblicate photoheterotrophic mutant of the cyanobacterium Synechocystis 6803. Inactivation of the psbA gene family. Plant Physiol 85: 1021–1025 (1987).

    Google Scholar 

  14. Kallio T, Aro E-M, Jansson C, Mäenpää P: Light-dependent turnover rate of the D1 polypeptide in new sitespecific Synechocystis PCC 6803 mutants. In: Proceedings of the 9th International Congress on Photosynthesis (in press).

  15. Kirilovsky DL, Ajlani G, Picaud M, Etienne A-L: Mutations responsible for high light sensitivity in an atrazineresistant mutant of Synechocystis 6714. Plant Mol Biol 13: 355–363 (1989).

    Google Scholar 

  16. Kirilovsky DL, Ducruet J-M, Etienne A-L: Primary events occurring in photoinhibition in Synechocystis 6714 wild-type and an atrazine-resistant mutant. Biochim Biophys Acta 1020: 87–93 (1990).

    Google Scholar 

  17. Krause GH, Weis E: Chlorophyll fluorescence and photosynthesis: the basics. Annu Rev Plant Physiol Plant Mol Biol 42: 313–349 (1991).

    Google Scholar 

  18. Krupa Z, Öquist G, Gustafsson P: Photoinhibition and recovery of photosynthesis in psbA gene-inactivated strains of cyanobacterium Anacystis nidulans. Plant Physiol 93: 1–6 (1990).

    Google Scholar 

  19. Kunkel TA, Roberts JD, Zakour RA: Rapid and efficient site-specific mutagenesis without phenotypic selection. Meth Enzymol 154: 367–382 (1987).

    Google Scholar 

  20. 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 

  21. Mackinney G: Absorption of light by chlorophyll solutions. J Biol Chem 140: 315–322 (1941).

    Google Scholar 

  22. Mattoo AK, Marder JB, Edelman M: Dynamics of the photosystem II reaction center. Cell 56: 241–246 (1989).

    Google Scholar 

  23. Michel H, Deisenhofer J: X-ray diffraction studies on a crystalline bacterial photosynthetic reaction center: a progress report and conclusions on the structure of photosystem II reaction center. In: Staehelin LA, Arntzen CJ (eds) Encyclopedia of Plant Physiology, vol 19, pp. 371–381. Springer-Verlag, Berlin (1986).

    Google Scholar 

  24. Miller AG, Sepie GS, Canvin DT: The effects of inorganic carbon and oxygen on fluorescence in the cyanobacterium Synechococcus UTEX 625. Can J Bot 69: 1151–1160 (1991).

    Google Scholar 

  25. Mohamed A, Eriksson J, Osiewacz HD, Jansson C: Differential expression of the psbA genes in the cyanobacterium Synechocystis 6803. Mol Gen Genet (in press).

  26. Mohamed A, Jansson C: Influence of light on accumulation of photosynthesis-specific transcripts in the cyanobacterium Synechocystis 6803. Plant Mol Biol 13: 693–700 (1989).

    Google Scholar 

  27. Ohad I, Koike H, Shochat S, Inoue Y: Changes in the properties of reaction center II during the initial stages of photoinhibition as revealed by thermoluminescence measurements. Biochim Biophys Acta 933: 288–298 (1988).

    Google Scholar 

  28. Ohad N, Amir-Shapira D, Koike H, Inoue Y, Ohad I, Hirschberg J: Amino acid substitutions in the D1 protein of photosystem II affect QB- stabilization and accelerate turnover of D1. Z Naturforsch 45c: 402–408 (1990).

    Google Scholar 

  29. Prasil O, Adir N, Ohad I: Dynamics of photosystem II: mechanism of photoinhibition and recovery processes. In: Barber J (ed) The Photosystems: Structure, Function and Molecular Biology. Topics in Photosynthesis, vol 11, pp. 295–348. Elsevier, Amsterdam (1992).

    Google Scholar 

  30. Prentki P, Kirsch HM: In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29: 303–313 (1984).

    Google Scholar 

  31. Rogers S, Wells R, Rechsteiner M: Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science 234: 364–368 (1986).

    Google Scholar 

  32. Salter AH, Virgin I, Hagman Å, Andersson B: On the molecular mechanism of light-induced D1 protein degradation in photosystem II core particles. Biochemistry 31: 3990–3998 (1992).

    Google Scholar 

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

    Google Scholar 

  34. Samuelsson G, Lönneborg A, Rosenqvist E, Gustafsson P, Öquist G. Photoinhibition and reactivation of photosynthesis in the cyanobacterium Anacystis nidulans. Plant Physiol 79: 992–995 (1985).

    Google Scholar 

  35. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5468 (1977).

    Google Scholar 

  36. Shipton CA, Marder JB, Barber J: Determination of catabolism of the photosystem II D1 subunit by structural motifs in the polypeptide sequence. Z Naturforsch 45c: 388–394 (1990).

    Google Scholar 

  37. Shipton CA, Barber J: Photoinduced degradation of the D1 polypeptide in isolated reaction centers of photosystem II: Evidence for an autoproteolytic process triggered by the oxidizing side of the photosystem. Proc Natl Acad Sci USA 88: 6691–6695 (1991).

    Google Scholar 

  38. Shochat S, Adir N, Gal A, Inoue Y, Mets L, Ohad I: Photoinactivation of photosystem II and degradation of the D1 protein are reduced in a cytochrome b6/f-less mutant of Chlamydomonas reinhardtii. Z Naturforsch 45c: 395–401 (1990).

    Google Scholar 

  39. Svensson B, Vass I, Cedergren E, Styring S: Structure of donor side components in photosystem II predicted by computer modelling. EMBO J 9: 2051–2059 (1990).

    Google Scholar 

  40. Svensson B, Vass I, Styring S: Sequence analysis of the D1 and D2 reaction center proteins of photosystem II. Z Naturforsch 46c: 765–776 (1991).

    Google Scholar 

  41. Trebst A: The topology of plastoquinone and herbicide binding peptides of photosystem II in the thylakoid membrane. Z Naturforsch 41c: 240–245 (1986).

    Google Scholar 

  42. Trebst A: A contact site between the two reaction center polypeptides of photosystem II is involved in photoinhibition. Z Naturforsch 46c: 557–562 (1991).

    Google Scholar 

  43. Trebst A, Depka B: Degradation of the D-1 protein subunit of photosystem II in isolated thylakoids by UV light. Z Naturforsch 45c: 765–771 (1990).

    Google Scholar 

  44. Trebst A, Depka B, Kipper M: The topology of the reaction center polypeptides of photosystem II. In: Baltscheffsky M (ed) Current Research in Photosynthesis, pp. 217–222. Kluwer Academic Publishers, Dordrecht (1990).

    Google Scholar 

  45. Trebst A, Draber W: Inhibitors of photosystem II and the topology of the herbicide and QB binding polypeptide in the thylakoid membrane. Photosynth Res 10: 381–382 (1986).

    Google Scholar 

  46. Tyystjärvi E, Karunen J: A microcomputer program and fast analog to digital converter and for the analysis of fluorescence induction transients. Photosynth Res 26: 127–132 (1990).

    Google Scholar 

  47. van der Bolt F, Vermaas W: Photoinactivation of Photosystem II as studied with site-directed mutants of the cyanobacterium Synechocystis sp. PCC6803. Biochim Biophys Acta 1098: 247–254 (1992).

    Google Scholar 

  48. Vass I, Styring S, Hundal T, Koivuniemi A, Aro E-M, Andersson B: Reversible and irreversible intermediates during photoinhibition of photosystem II: Stable reduced QA species promote chlorophyll triplet formation. Proc Natl Acad Sci USA 89: 1408–1412 (1992).

    Google Scholar 

  49. Virgin I, Ghanotakis DF, Andersson B: Light-induced D1-protein degradation in isolated photosystem II core complexes. FEBS Lett 269: 45–48 (1990).

    Google Scholar 

  50. 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 

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Authors and Affiliations

  1. Dept. of Biology, University of Turku, SF-20700, Turku, Finland

    Pirkko Mäenpää, Taina Kallio, Paula Mulo, Eva-Mari Aro & Esa Tyystjärvi

  2. Dept. of Biochemistry, University of Stockholm, S-10691, Stockholm, Sweden

    Gaza Salih & Christer Jansson

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  1. Pirkko Mäenpää
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  2. Taina Kallio
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  3. Paula Mulo
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  4. Gaza Salih
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  7. Christer Jansson
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Mäenpää, P., Kallio, T., Mulo, P. et al. Site-specific mutations in the D1 polypeptide affect the susceptibility of Synechocystis 6803 cells to photoinhibition. Plant Mol Biol 22, 1–12 (1993). https://doi.org/10.1007/BF00038991

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