Abstract
The degradation rate of the D1 polypeptide was measured in threeSynechocystis PCC 6803 mutantsin vivo. Mutations were introduced into a putative cleavage area of the D1 polypeptide (QEEET motif) and into the PEST-like area. PEST sequences are often found in proteins with a high turnover rate. The QEEET-motif mutants are CA1 [Δ(E242-E244);Q241H] and E243K, and the third mutation, E229D, was directed to the PEST-like area. During high-light illumination (1500 μmol photons m-2s-1) that induced photoinhibition of photosystem II (PSII), the half-life time of the D1 polypeptide in mutant E229D (t 1/2=35 min) was about twice as long as in AR (control strain) cells (t 1/2=19 min). In growth light (40 μmol photons m-2s-1), the degradation rate of the D1 polypeptide in E229D and AR strains was the same (t 1/2≈5 h). In growth light the D1 polypeptide was degraded faster in both QEEET-motif mutants than in the AR strain, but in photoinhibitory light the degradation rates were similar. According to these results, the highly conservative QEEET motif as such is not required for the proteolytic cut of the D1 polypeptide, but it does affect the rate of degradation. No simple correlation existed between the degradation rate of the D1 polypeptide and the susceptibility of PSII to photoinhibition in mutant and AR cells under our experimental conditions.
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References
Andersson B, Styring S: Photosystem II: Molecular organization, function, and acclimation. Curr Topics Bioenerget 16: 1–81 (1991).
Arnon DI: Copper enzymes in isolated chloroplasts. Polyphenoloxidase inBeta vulgaris. Plant Physiol 24: 1–15 (1949).
Aro E-M, Hundal T, Carlberg I and Andersson B:In vitro studies on light-induced inhibition of Photosystem II and D1-protein degradation at low temperatures. Biochim Biophys Acta 1019: 262–275 (1990).
Barbato R, Shipton CA, Giacometti GM, Barber J: New evidence suggests that the initial photoinduced cleavage of the D1-protein may not occur near the PEST sequence. FEBS Lett 290: 162–166 (1991).
Bennet A, Bogorad L. Complementary chromatic adaptation in a filamentous blue-green alga. J Cell Biol 58: 419–435 (1973).
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).
Erickson JM, Rochaix J-D: The molecular biology of Photosystem II. In: Barber J (ed) The Photosystems: Structure, Function and Molecular Biology. Topics in Photosynthesis, vol 11, pp. 101–177, Elsevier, Amsterdam (1992).
Greenberg BM, Gaba V, Canaani O, Malkin S, Mattoo AK, Edelman M: Separate photosensitizers mediate degradation of the 32-kDa photosystem II reaction centre protein in the visible and UV spectral regions. Proc Natl Acad Sci USA 86: 6617–6620 (1989).
Greenberg BM, Gaba V, Mattoo AK, and Edelman M: Identification of a primary in vivo degradation product of the rapidly-turning-over 32 kd protein of photosystem II. EMBO J 6: 2865–2869 (1987).
Kanervo E, Mäenpää P, Aro E-M: D1 protein degradation andpsbA transcript levels inSynechocystis PCC 6803 during photoinhibitionin vivo. J Plant Physiol, in press (1994).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).
Mattoo AK, Hoffman-Falk H, Marder JB, Edelman M: Regulation of protein metabolism: coupling of photosynthetic electron transport toin vivo degradation of the rapidly metabolized 32-kilodalton protein of the chloroplast membranes. Proc Natl Acad Sci USA 81: 1380–1384 (1984).
Misra AN, Hall SG, Barber J: The isolated D1/D2/cyt b-559 reaction centre complex of Photosystem II possesses a serine-type endopeptidase activity. Biochim Biophys Acta 1059: 239–242 (1991).
Mohamed A, Jansson C: Influence of light on accumulation of photosynthesis-specific transcripts in the cyanobacteriumSynechocystis 6803. Plant Mol Biol 1: 693–700 (1989).
Mäenpää P, Kallio T, Mulo P, Salih G, Aro E-M, Tyystjärvi E, Jansson C: Site-specific mutations in the D1 polypeptide affect the susceptibility ofSynechocystis 6803 cells to photoinhibition. Plant Mol Biol 22: 1–12 (1993).
Nanba O, Satoh K: Isolation of a photosystem II reaction centre consisting of D-1 and D-2 polypeptides and cytochrome b-559. Proc Natl Acad Sci USA 84: 109–112 (1987).
Ohad I, Kyle DJ, Hirschberg J: Light-dependent degradation of the QB-protein in isolated pea thylakoids. EMBO J 4: 1655–1659 (1985).
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).
Ohad N, Inoue Y, Hirschberg J: Accelerated rate of turnover of the D1 subunit of photosystem II is correlated with inhibition of electron transfer from QA to QB in cyanobacterial mutants. In: Argyroudi-Akoyunologlou JH (ed) Regulation of Chloroplast Biogenesis, pp. 589–596, Plenum Press, New York (1992).
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, Amsterdam, Elsevier (1992).
Rogers S, Wells R, Rechsteiner M: Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science 234: 364–368 (1986).
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).
Shipton CA, Marder JB and Barber J: Determination of catabolism of the photosystem II D1 subunit by structural motifs in the polypeptide sequence Z Naturforsch 45c: 388–394 (1989).
Tae GS, Cramer WA: Truncation of the COOH-terminal domain of the psbE gene product inSynechocystis sp. PCC 6803: requirements for photosystem II assembly and function. Biochemistry 31: 4066–4074 (1992).
Trebst A. The topology of plastochinon and herbicide binding peptides of Photosystem II in the thylakoid membrane. Z Naturforsch 41c: 240–245 (1986).
Trebst A, Depka B: Dégradation of the D-1 protein subunit of photosystem II in isolated thylakoids by UV light. Z Naturforsch 45c: 765–771 (1990).
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).
Virgin I, Salter AH, Ghanotakis DF, Andersson B: Light-induced D1 protein degradation is catalyzed by a serinetype protease. FEBS Lett 287: 125–128 (1991).
Zurawski G, Bohnert HH, Whitfeld PR, Bottomley W: Nucleotide sequence of the gene for theM r 32,000 thylakoid membrane protein fromSpinacia oleracea andNicotiana debneyi predicts a totally conserved primary translation product ofM r 38,950. Proc Natl Acad Sci USA 79: 7699–7703 (1982).
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Tyystjärvi, T., Aro, EM., Jansson, C. et al. Changes of amino acid sequence in PEST-like area and QEEET motif affect degradation rate of D1 polypeptide in photosystem II. Plant Mol Biol 25, 517–526 (1994). https://doi.org/10.1007/BF00043879
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DOI: https://doi.org/10.1007/BF00043879