Skip to main content
SpringerLink
Log in

Light activates binding of membrane proteins to chloroplast RNAs in Chlamydomonas reinhardtii

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Several membrane proteins were previously shown to bind to the 5′ leader of the chloroplast psbC mRNA in the unicellular eukaryotic alga Chlamydomonas reinhardtii. This study showed that these proteins have affinity for AU-rich RNAs, as determined by competition experiments. In addition, their binding activities are enhanced 13–15-fold by light, and a 46 kDa protein is activated within 1–10 min. This activation could be mediated by the modulation of ADP pools by the light-dependent reactions of photosynthesis and ATP synthase because (1) two inhibitors that block ATP synthesis also prevent this activation and (2) ADP inhibits the RNA-binding activity of this protein in vitro. An inhibitor of Photosystem II diminishes this induction, suggesting that reducing potential generated by the photosynthetic electron transport chain modulates this RNA-binding activity. The RNA-binding activities of two proteins (of 46 and 47 kDa) are inhibited by Mg-protoporphyrin IX methyl ester in vitro suggesting they could be regulated by these intermediates in the chlorophyll biosynthetic pathway.

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

  • Adam, Z. 2000. Chloroplast proteases: possible regulators of gene expression? Biochimie 82: 647–654.

    Google Scholar 

  • Allen, J.F. and Raven, J.A. 1996. Free-radical-induced mutation vs redox regulation: costs and benefits of genes in organelles. J. Mol. Evol. 42: 482–492.

    Google Scholar 

  • Barkan, A. and Goldschmidt-Clermont, M. 2000. Participation of nuclear genes in chloroplast gene expression. Biochimie 82: 559–572.

    Google Scholar 

  • Bennoun, P. and Beal, D 1997. Screening algal mutant colonies with altered thylakoid electronchemical gradient through fluorescence and delayed luminescence digital imaging. Photosynth. Res. 51: 161–165.

    Google Scholar 

  • Bruick, R.K. and Mayfield, S.P. 1998. Processing of the psbA 5' untranslated region in Chlamydomonas reinhardtii depends upon factors mediating ribosome association. J. Cell Biol. 143: 1145–1153.

    Google Scholar 

  • Bruick, R.K. and Mayfield, S.P. 1999. Light-activated translation of chloroplast mRNAs. Trends Plant Sci. 4 (5): 190–195.

    Google Scholar 

  • Bulte, L. and Bennoun, P. 1990. Translational accuracy and sexual differentiation in Chlamydomonas reinhardtii. Curr. Genet. 18: 155–160.

    Google Scholar 

  • Bulte, L. and Wollman, F.A. 1992. Evidence for a selective destabilization of an integral membrane protein, the cytochrome b6/f complex, during gametogenesis in Chlamydomonas reinhardtii. Eur. J. Biochem. 204: 327–336.

    Google Scholar 

  • Caliebe, A. and Soill, J. 1999. News in chloroplast protein import. Plant Mol Biol. 39: 641–645.

    Google Scholar 

  • Choquet, Y. and Vallon, O. 2000. Synthesis, assembly and degradation of thylakoid membrane proteins. Biochimie 82: 615–634.

    Google Scholar 

  • Christopher, D.A., Li, X., Kim, M. and Mullet, J.E. 1997. Involvement of protein kinase and extraplastidic serine/threonine protein phosphatases in signaling pathways regulating plastid transcription and the psbD blue light-responsive promoter in barley. Plant Physiol. 113: 1273–1282.

    Google Scholar 

  • Cohen, A. and Mayfield, S.P. 1997. Translational regulation of gene expression in plants. Curr. Opin. Biotechnol. 8: 189–194.

    Google Scholar 

  • Danon, A. and Mayfield, S.P. 1991. Light regulated translational activators: identification of chloroplast gene specific mRNA binding proteins. EMBO J. 10: 3993–4001.

    Google Scholar 

  • Danon, A. and Mayfield, S.P. 1994a. ADP-dependent phosphorylation regulates RNA-binding in vitro: implications in lightmodulated translation. EMBO J. 13: 2227–2235.

    Google Scholar 

  • Danon, A. and Mayfield, S.P. 1994b. Light-regulated translation of chloroplast messenger RNAs through redox potential. Science 266: 1717–1719.

    Google Scholar 

  • Davies, D.R., and Plaskitt, A 1971. Genetical and structural analyses of cell-wall formation in Chlamydomonas reinhardtii. Genet. Res. 17: 33–43.

    Google Scholar 

  • Detmers, P.A., Goodenough, U.W. and Condeelis, J. 1983. Elongation of the fertilization tubule in Chlamydomonas: new observations on the core microfilaments and the effect of transient intracellular signals on their structural integrity. J. Cell Biol. 97: 522–532.

    Google Scholar 

  • Escoubas, J.M., Lomas, M., LaRoche, J. and Falkowski, P.G. 1995. Light intensity regulation of cab gene transcription is signaled by the redox state of the plastoquinone pool. Proc. Natl. Acad. Sci. USA 92: 10237–10241.

    Google Scholar 

  • Fargo, D., Boynton, J. and Gillham, N. 2001. Chloroplast ribosomal protein S7 of Chlamydomonas binds to chloroplast mRNA leader sequences and may be involved in translation initiation. Plant Cell 13: 207–218.

    Google Scholar 

  • Gorman, D.S. and Levine, R.P. 1965. Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardtii. Proc. Natl. Acad. Sci. USA 54: 1665–1669.

    Google Scholar 

  • Gray, M.W. 1999. Evolution of organellar genomes. Curr. Opin. Genet. Dev. 9: 678–687.

    Google Scholar 

  • Hauser, C.R., Gillham, N.W. and Boynton, J.E. 1996. Translational regulation of chloroplast genes. Proteins binding to the 5'-untranslated regions of chloroplast mRNAs in Chlamydomonas reinhardtii. J. Biol. Chem. 271: 1486–1497.

    Google Scholar 

  • Hauser, C.R., Gillham, N.W., and Boynton, J.E. 1998. Regulation of chloroplast translation. In: G.-C. M., J.-D. Rochaix and S. Merchant (Eds.) The Molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas, Kluwer Academic Publishers, Norwell, MA/Dordrecht, Netherlands, pp. 197–217.

    Google Scholar 

  • Hoober, R.A.W.J.K. 1994. Biogenesis of thylakoid membranes in Chlamydomonas reinhardtii y1. Plant Physiol. 106: 583–590.

    Google Scholar 

  • Hwang, S., Kawazoe, R. and Herrin, D.L. 1996. Transcription of tufA and other chloroplast-encoded genes is controlled by a circadian clock in Chlamydomonas. Proc. Natl. Acad. Sci. USA 93: 996–1000.

    Google Scholar 

  • Inada, H., Kusumi, K., Nishimura, M. and Iba, K. 1996. Specific expression of the chloroplast gene for RNA polymerase (rpoB) at an early stage of leaf development in rice. Plant Cell Physiol. 37: 229–232.

    Google Scholar 

  • Irihimovitch, V. and Shapira, M. 2000. Glutathione redox potential modulated by reactive oxygen species regulates translation of Rubisco large subunit in the chloroplast. J. Biol. Chem. 275: 16289–16295.

    Google Scholar 

  • Jarvis, P. 2001. Intracellular signaling: the chloroplast talks! Curr. Biol. 11: R307–R310.

    Google Scholar 

  • Johanningmeier, U. 1988. Possible control of transcript levels by chlorophyll precursors in Chlamydomonas. Eur. J. Biochem. 177: 417–424.

    Google Scholar 

  • Johanningmeier, U. and Howell, S.H. 1984. Regulation of lightharvesting chlorophyll-binding protein mRNA accumulation in Chlamydomonas reinhardtii. Possible involvement of chlorophyll synthesis precursors. J. Biol. Chem. 259: 13541–13549.

    Google Scholar 

  • Joyard, J., Block, M. A., and Douce, R. 1991. Molecular aspects of plastid envelope biochemistry. Eur. J. Biochem. 199: 489–509.

    Google Scholar 

  • Kannangara, C.G., Vothknecht, U.C., Hansson, M. and von Wettstein, D. 1997. Magnesium chelatase: association with ribosomes and mutant complementation studies identify barley subunit Xantha-G as a functional counterpart of Rhodobacter subunit BchD. Mol. Gen. Genet. 254: 85–92.

    Google Scholar 

  • Kim, J. and Mayfield, S.P. 1997. Protein disulfide isomerase as a regulator of chloroplast translational activation. Science 278: 1954–1957.

    Google Scholar 

  • Kropat, J., Oster, U., Rudiger, W. and Beck, C.F. 1997. Chlorophyll precursors are signals of chloroplast origin involved in light induction of nuclear heat-shock genes. Proc. Natl. Acad. Sci. USA 94: 14168–14172.

    Google Scholar 

  • Kropat, J., Oster, U., Rudiger, W. and Beck, C.F. 2000. Chloroplast signalling in the light induction of nuclear HSP70 genes requires the accumulation of chlorophyll precursors and their accessibility to cytoplasm/nucleus. Plant J. 24: 523–531.

    Google Scholar 

  • Malnoe, P., Mayfield, S.P. and Rochaix, J.D. 1988. Comparative analysis of the biogenesis of Photosystem II in the wild-type and Y-1 mutant of Chlamydomonas reinhardtii. J. Cell Biol. 106: 609–616.

    Google Scholar 

  • Martin, N.C. and Goodenough, U.W. 1975. Gametic differentiation in Chlamydomonas reinhardtii. I. Production of gametes and their fine structure. J. Cell Biol. 67: 587–605.

    Google Scholar 

  • Mayfield, S.P. 1990. Chloroplast gene regulation: interaction of the nuclear and chloroplast genomes in the expression of photosynthetic proteins. Curr. Opin. Cell Biol. 2: 509–513.

    Google Scholar 

  • Mayfield, S.P., Cohen, A., Danon, A. and Yohn, C.B. 1994. Translation of the psbA mRNA of Chlamydomonas reinhardtii requires a structured RNA element contained within the 5' untranslated region. J. Cell Biol. 127: 1537–1545.

    Google Scholar 

  • Mochizuki, N., Brusslan, J.A., Larkin, R., Nagatani, A. and Chory, J. 2001. Arabidopsis genomes uncoupled 5 (GUN5) mutant reveals the involvement of Mg-chelatase H subunit in plastidto-nucleus signal transduction. Proc. Natl. Acad. Sci. USA 98: 2053–2058.

    Google Scholar 

  • Moller, S.G., Kunkel, T. and Chua, N.H. 2001. A plastidic ABC protein involved in intercompartmental communication of light signaling. Genes Dev. 15: 90–103.

    Google Scholar 

  • Monde, R.A., Schuster, G. and Stern, D.B. 2000. Processing and degradation of chloroplast mRNA. Biochimie 82: 573–582.

    Google Scholar 

  • Muhlbauer, S.K. and Eichacker, L.A. 1998. Light-dependent formation of the photosynthetic proton gradient regulates translation elongation in chloroplasts. J. Biol. Chem. 273: 20935–20940.

    Google Scholar 

  • Mullet, J.E. 1993. Dynamic regulation of chloroplast transcription. Plant Physiol. 103: 309–313.

    Google Scholar 

  • Nickelsen, J. 1998. Chloroplast mRNA stability. In: M.G.-C. (AUTHOR: PLEAE GIVE FULL NAME), J.-D. Rochaix and S. Merchant (Eds) The Molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas, Kluwer Academic Publishers, Norwell, MA/Dordrecht, Netherlands, pp. 151–163.

    Google Scholar 

  • Nickelsen, J., van Dillewijn, J., Rahire, M. and Rochaix, J.D. 1994. Determinants for stability of the chloroplast psbD RNA are located within its short leader region in Chlamydomonas reinhardtii. EMBO J. 13: 3182–3191.

    Google Scholar 

  • Pfannschmidt, T., Nilsson, A., Tullberg, A., Link, G. and Allen, J.F. 1999. Direct transcriptional control of the chloroplast genes psbA and psaB adjusts photosynthesis to light energy distribution in plants. IUBMB Life 48: 271–276.

    Google Scholar 

  • Popperl, G., Oster, U. and Rudiger, W. 1998. Light-dependent increase in chlorophyll precursors during the day-night cycle in tobacco and barley seedlings. J. Plant Physiol. 153: 40–45.

    Google Scholar 

  • Rochaix, J.D. 1992. Post-transcriptional steps in the expression of chloroplast genes. Annu. Rev. Cell Biol. 8: 1–28.

    Google Scholar 

  • Rochaix, J.D. 1996. Post-transcriptional regulation of chloroplast gene expression in Chlamydomonas reinhardtii. Plant Mol. Biol. 32: 327–341.

    Google Scholar 

  • Rott, R., Levy, H., Drager, R.G., Stern, D.B. and Schuster, G. 1998. 3'-Processed mRNA is preferentially translated in Chlamydomonas reinhardtii chloroplasts. Mol. Cell Biol. 18: 4605–4611.

    Google Scholar 

  • Sambrook, J., Fritsch, E. F., and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.

    Google Scholar 

  • Sato, N., Rolland, N., Block, M.A. and Joyard, J. 1999. Do plastid envelope membranes play a role in the expression of the plastid genome? Biochimie 81: 619–629.

    Google Scholar 

  • Shapira, M., Lers, A., Heifetz, P.B., Irihimovitz, V., Osmond, C.B., Gillham, N.W. and Boynton, J.E. 1997. Differential regulation of chloroplast gene expression in Chlamydomonas reinhardtii during photoacclimation: light stress transiently suppresses synthesis of the Rubisco LSU protein while enhancing synthesis of the PS II D1 protein. Plant Mol. Biol. 33: 1001–1011.

    Google Scholar 

  • Shochat, S., Owens, G. C., Hubert, P. and Ohad, I. 1982. The dichlorophenyldimethyl-urea-binding site in thylakoids of Chlamydomonas reinhardtii. Role of Photosystem II reaction center and phosphorylation of the 32-35 kilodalton polypeptide in the formation of the high-affinity binding site. Biochim. Biophys. Acta 681: 21–31.

    Google Scholar 

  • Smith, H.B. 2000. Photoreceptors in signal transduction. Pathways of enlightenment. Plant Cell 12: 1–3.

    Google Scholar 

  • Smith, P.K., Krohn, R.I., Hermanson, G.T., Mallia, A.K., Gartner, F.H., Provenzano, M.D., Fujimoto, E.K., Goeke, N.M., Olson, B.J. and Klenk, D.C. 1985. Measurement of protein using bicinchoninic acid. Anal. Biochem. 150: 76–85.

    Google Scholar 

  • Staub, J.M. and Maliga, P. 1993. Accumulation of D1 polypeptide in tobacco plastids is regulated via the untranslated region of the psbA mRNA. EMBO J. 12: 601–606.

    Google Scholar 

  • Sugiura, M., Hirose, T. and Sugita, M. 1998. Evolution and mechanism of translation in chloroplasts. Annu. Rev. Genet. 32: 437–459.

    Google Scholar 

  • Takahashi, Y., Utsumi, K., Yamamoto, Y., Hatano, A. and Satoh, K. 1996. Genetic engineering of the processing site of D1 precursor protein of Photosystem II reaction center in Chlamydomonas reinhardtii. Plant Cell Physiol. 37: 161–168.

    Google Scholar 

  • Trebitsh, T. and Danon, A. 2001. Translation of chloroplast psbA mRNA is regulated by signals initiated by both Photosystems II and I. Proc. Natl. Acad. Sci. USA 98: 12289–12294.

    Google Scholar 

  • Trebitsh, T., Meiri, E., Ostersetzer, O., Adam, Z. and Danon, A. 2001. The protein disulfide isomerase-like RB60 is partitioned between stroma and thylakoids in Chlamydomonas reinhardtii chloroplasts. J. Biol. Chem. 276: 4564–4569.

    Google Scholar 

  • von Gromoff, E.D. and Beck, C.F. 1993. Genes expressed during sexual differentiation of Chlamydomonas reinhardtii. Mol. Gen. Genet. 241: 415–421.

    Google Scholar 

  • Yohn, C.B., Cohen, A., Danon, A. and Mayfield, S.P. 1996. Altered mRNA binding activity and decreased translational initiation in a nuclear mutant lacking translation of the chloroplast psbA mRNA. Mol. Cell Biol. 16: 3560–3566.

    Google Scholar 

  • Yohn, C.B., Cohen, A., Danon, A. and Mayfield, S.P. 1998a. A poly(A) binding protein functions in the chloroplast as a message-specific translation factor. Proc. Natl. Acad. Sci. USA 95: 2238–2243.

    Google Scholar 

  • Yohn, C.B., Cohen, A., Rosch, C., Kuchka, M.R. and Mayfield, S.P. 1998b. Translation of the chloroplast psbA mRNA requires the nuclear-encoded poly(A)-binding protein, RB47. J. Cell Biol. 142: 435–442.

    Google Scholar 

  • Zerges, W. 2000. Translation in chloroplasts. Biochimie 82: 583–601.

    Google Scholar 

  • Zerges, W. 2002. Does complexity constrain organelle evolution? Trends Plant Sci. 7 (4): 175–182.

    Google Scholar 

  • Zerges, W. and Rochaix, J.D. 1994. The 5' leader of a chloroplast mRNA mediates the translational requirements for two nucleus-encoded functions in Chlamydomonas reinhardtii. Mol. Cell Biol. 14: 5268–5277.

    Google Scholar 

  • Zerges, W. and Rochaix, J.D. 1998. Low density membranes are associated with RNA-binding proteins and thylakoids in the chloroplast of Chlamydomonas reinhardtii. J. Cell Biol. 140: 101–110.

    Google Scholar 

  • Zhang, L., Paakkarinen, V., van Wijk, K.J. and Aro, E.M. 2000. Biogenesis of the chloroplast-encoded D1 protein: regulation of translation elongation, insertion, and assembly into Photosystem II. Plant Cell 12: 1769–1782.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biology Department, Concordia University, 1445 Maisonneuve W., Montreal, Canada, H3G 1M8

    William Zerges & Shengwu Wang

  2. Departments of Molecular Biology and Plant Biology, University of Geneva, 30 Quai Ernest-Ansermet, 1211, Geneva 4, Switzerland

    Jean-David Rochaix

Authors
  1. William Zerges
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shengwu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jean-David 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

Zerges, W., Wang, S. & Rochaix, JD. Light activates binding of membrane proteins to chloroplast RNAs in Chlamydomonas reinhardtii . Plant Mol Biol 50, 573–585 (2002). https://doi.org/10.1023/A:1020246007858

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1020246007858

Search

Navigation