Carbon Dioxide; Photosystem II Protein Complex; Proteins; Photosystem II Protein Complex/metabolism; Photosynthesis/genetics; Proteins/metabolism; Carbon Dioxide/metabolism; Chlamydomonas reinhardtii/metabolism; Chlamydomonas reinhardtii; Photosynthesis; Chemistry (all); Biochemistry, Genetics and Molecular Biology (all); Physics and Astronomy (all); General Physics and Astronomy; General Biochemistry, Genetics and Molecular Biology; General Chemistry; Multidisciplinary
Abstract :
[en] Photosynthetic algae have evolved mechanisms to cope with suboptimal light and CO2 conditions. When light energy exceeds CO2 fixation capacity, Chlamydomonas reinhardtii activates photoprotection, mediated by LHCSR1/3 and PSBS, and the CO2 Concentrating Mechanism (CCM). How light and CO2 signals converge to regulate these processes remains unclear. Here, we show that excess light activates photoprotection- and CCM-related genes by altering intracellular CO2 concentrations and that depletion of CO2 drives these responses, even in total darkness. High CO2 levels, derived from respiration or impaired photosynthetic fixation, repress LHCSR3/CCM genes while stabilizing the LHCSR1 protein. Finally, we show that the CCM regulator CIA5 also regulates photoprotection, controlling LHCSR3 and PSBS transcript accumulation while inhibiting LHCSR1 protein accumulation. This work has allowed us to dissect the effect of CO2 and light on CCM and photoprotection, demonstrating that light often indirectly affects these processes by impacting intracellular CO2 levels.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Águila Ruiz-Sola, M ; Univ. Grenoble Alpes, CNRS, CEA, INRAE, IRIG-LPCV, 38000, Grenoble, France ; Instituto de Bioquímica Vegetal y Fotosíntesis, Consejo Superior de Investigaciones Científicas-Universidad de Sevilla, Sevilla, Spain
Sanz-Luque, Emanuel ; The Carnegie Institution for Science, Department of Plant Biology, Stanford, CA, 94305, USA ; University of Cordoba, Department of Biochemistry and Molecular Biology, Cordoba, Spain
Redekop, Petra ; The Carnegie Institution for Science, Department of Plant Biology, Stanford, CA, 94305, USA
Tokutsu, Ryutaro ; Division of Environmental photobiology, National Institute for Basic Biology (NIBB), Nishigonaka 38, Myodaiji, Okazaki, 444-8585, Japan
Küken, Anika ; Bioinformatics Group, Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany ; Max-Planck-Institute of Molecular Plant Physiology, Potsdam, Golm, Germany
Arend, Marius ; Bioinformatics Group, Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany ; Max-Planck-Institute of Molecular Plant Physiology, Potsdam, Golm, Germany
Finazzi, Giovanni ; Univ. Grenoble Alpes, CNRS, CEA, INRAE, IRIG-LPCV, 38000, Grenoble, France
Hippler, Michael ; Institute of Plant Biology and Biotechnology, Westfälische Wilhelms Universität, 48143, Münster, Germany
Nikoloski, Zoran; Bioinformatics Group, Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany ; Max-Planck-Institute of Molecular Plant Physiology, Potsdam, Golm, Germany
Minagawa, Jun ; Division of Environmental photobiology, National Institute for Basic Biology (NIBB), Nishigonaka 38, Myodaiji, Okazaki, 444-8585, Japan
Grossman, Arthur R ; The Carnegie Institution for Science, Department of Plant Biology, Stanford, CA, 94305, USA
We are grateful to Dr. Konomi Fujimura-Kamada for performing experiments to validate the dark induction of LHCSR3 in the Minagawa lab; to Claire Remacle for sharing the icl and icl-C strains and Pierre Cardol for the dum11 strain. We thank Eric Soupene for valuable insights in the RHP1 induction conditions. We thank Dimitra Karageorgou for performing preliminary experiments in the project and Gilles Curien for fruitful discussions on aspects of microalgae metabolism. We would like to thank the following agencies for funding: The Human Frontiers Science Program through the funding of the project RGP0046/2018 (DP, ARG, PR, ES-L, ZN, AK); the French National Research Agency in the framework of the Young Investigators program ANR-18-CE20-0006 through the funding of the project MetaboLight (DP); the French National Research Agency through the funding of the Grenoble Alliance for Integrated Structural & Cell Biology GRAL project ANR-17-EURE-0003 (DP, MAR-S, GV, YY); the French National Research Agency in the framework of the Investissements d’Avenir program ANR-15-IDEX-02, through the funding of the “Origin of Life” project of the Univ. Grenoble-Alpes (DP, YY); the Prestige Marie-Curie co-financing grant PRESTIGE-2017-1-0028 (MAR-S); the International Max Planck Research School “Primary Metabolism and Plant Growth” at the Max Planck Institute of Molecular Plant Physiology (MA, ZN); the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement no. 751039 (ES-L); the program ‘Plan Propio UCO’ from University of Cordoba, Spain for postdoctoral Support (ES-L); the Carnegie Institution for Science and the Department of Energy, DE-SC0019417 (ARG); the Marie Curie Initial Training Network Accliphot FP7-PEPOPLE-2012-ITN; 316427 (SF, GF, DP); the Japan Society for the Promotion of Science, JSPS, for the grants-in-Aid for Scientific Research, KAKENHI, 21H04778 and 21H05040 (JM) and the German Research Foundation DFG HI 739/9.2 (MH).
Li, Z., Wakao, S., Fischer, B. B. & Niyogi, K. K. Sensing and responding to excess light. Annu. Rev. Plant Biol. 60, 239–260 (2009). DOI: 10.1146/annurev.arplant.58.032806.103844
Bennett, D. I. G. et al. Models and mechanisms of the rapidly reversible regulation of photosynthetic light harvesting. Open Biol. 9, 190043 (2019). DOI: 10.1098/rsob.190043
Alboresi, A., Gerotto, C., Giacometti, G. M., Bassi, R. & Morosinotto, T. Physcomitrella patens mutants affected on heat dissipation clarify the evolution of photoprotection mechanisms upon land colonization. Proc. Natl Acad. Sci. USA. 107, 11128–11133 (2010). DOI: 10.1073/pnas.1002873107
Niyogi, K. K. & Truong, T. B. Evolution of flexible non-photochemical quenching mechanisms that regulate light harvesting in oxygenic photosynthesis. Curr. Opin. Plant Biol. 16, 307–314 (2013). DOI: 10.1016/j.pbi.2013.03.011
Peers, G. et al. An ancient light-harvesting protein is critical for the regulation of algal photosynthesis. Nature 462, 518–521 (2009). DOI: 10.1038/nature08587
Correa-Galvis, V. et al. Photosystem II Subunit PsbS Is Involved in the Induction of LHCSR Protein-dependent Energy Dissipation in Chlamydomonas reinhardtii. J. Biol. Chem. 291, 17478–17487 (2016). DOI: 10.1074/jbc.M116.737312
Tibiletti, T., Auroy, P., Peltier, G. & Caffarri, S. Chlamydomonas reinhardtii PsbS Protein Is Functional and Accumulates Rapidly and Transiently under High Light. Plant Physiol. 171, 2717–2730 (2016). DOI: 10.1104/pp.16.00572
Allorent, G. et al. UV-B photoreceptor-mediated protection of the photosynthetic machinery in Chlamydomonas reinhardtii. Proc. Natl Acad. Sci. USA. 113, 14864–14869 (2016). DOI: 10.1073/pnas.1607695114
Dinc, E. et al. LHCSR1 induces a fast and reversible pH-dependent fluorescence quenching in LHCII in Chlamydomonas reinhardtii cells. Proc. Natl Acad. Sci. USA. 113, 7673–7678 (2016). DOI: 10.1073/pnas.1605380113
Petroutsos, D. et al. A blue-light photoreceptor mediates the feedback regulation of photosynthesis. Nature 537, 563–566 (2016). DOI: 10.1038/nature19358
Petroutsos, D. et al. The chloroplast calcium sensor CAS is required for photoacclimation in Chlamydomonas reinhardtii. Plant Cell 23, 2950–2963 (2011). DOI: 10.1105/tpc.111.087973
Tokutsu, R., Fujimura-Kamada, K., Matsuo, T., Yamasaki, T. & Minagawa, J. The CONSTANS flowering complex controls the protective response of photosynthesis in the green alga Chlamydomonas. Nat. Commun. 10, 655–10 (2019). DOI: 10.1038/s41467-019-11989-x
Gabilly, S. T. et al. Regulation of photoprotection gene expression in Chlamydomonas by a putative E3 ubiquitin ligase complex and a homolog of CONSTANS. Proc. Natl Acad. Sci. USA. 116, 17556–17562 (2019). DOI: 10.1073/pnas.1821689116
Maberly, S. C. & Gontero, B. Ecological imperatives for aquatic CO2-concentrating mechanisms. J. Exp. Bot. 68, 3797–3814 (2017). DOI: 10.1093/jxb/erx201
Moroney, J. V. et al. Isolation and Characterization of a Mutant of Chlamydomonas reinhardtii Deficient in the CO2 Concentrating Mechanism. Plant Physiol. 89, 897–903 (1989). DOI: 10.1104/pp.89.3.897
Fukuzawa, H. et al. Ccm1, a regulatory gene controlling the induction of a carbon-concentrating mechanism in Chlamydomonas reinhardtii by sensing CO2 availability. Proc. Natl Acad. Sci. USA. 98, 5347–5352 (2001). DOI: 10.1073/pnas.081593498
Xiang, Y., Zhang, J. & Weeks, D. P. The Cia5 gene controls formation of the carbon concentrating mechanism in Chlamydomonas reinhardtii. Proc. Natl Acad. Sci. USA. 98, 5341–5346 (2001). DOI: 10.1073/pnas.101534498
Harris, E. H. Chlamydomonas as a Model Organism. Annu. Rev. Plant. Physiol. 10.1146/annurev.arplant.52.1.363 (2001).
Wolfe, A. J. The acetate switch. Microbiol. Mol. Biol. Rev. 69, 12–50 (2005). DOI: 10.1128/MMBR.69.1.12-50.2005
Kornberg, H. L. & Krebs, H. A. Synthesis of cell constituents from C2-units by a modified tricarboxylic acid cycle. Nature 179, 988–991 (1957). Nature. DOI: 10.1038/179988a0
Plancke, C. et al. Lack of isocitrate lyase in Chlamydomonas leads to changes in carbon metabolism and in the response to oxidative stress under mixotrophic growth. Plant J. 77, 404–417 (2014). DOI: 10.1111/tpj.12392
Salinas, T., Larosa, V., Cardol, P., Marechal-Drouard, L. & Remacle, C. Respiratory-deficient mutants of the unicellular green alga Chlamydomonas: A review. Biochimie. https://doi.org/10.1016/j.biochi.2013.10.006 (2013).
Fett, J. P. & Coleman, J. R. Regulation of Periplasmic Carbonic Anhydrase Expression in Chlamydomonas reinhardtii by Acetate and pH. Plant Physiol. 106, 103–108 (1994). DOI: 10.1104/pp.106.1.103
Moroney, J. V., Kitayama, M., Togasaki, R. K. & Tolbert, N. E. Evidence for Inorganic Carbon Transport by Intact Chloroplasts of Chlamydomonas reinhardtii. Plant Physiol. 83, 460–463 (1987). DOI: 10.1104/pp.83.3.460
Polukhina, I., Fristedt, R., Dinc, E., Cardol, P. & Croce, R. Carbon supply and photoacclimation crosstalk in the green alga Chlamydomonas reinhardtii. Plant Physiol. 1494–1505. https://doi.org/10.1104/pp.16.01310 (2016)
Fang, W. et al. Transcriptome-wide changes in Chlamydomonas reinhardtii gene expression regulated by carbon dioxide and the CO2-concentrating mechanism regulator CIA5/CCM1. Plant Cell 24, 1876–1893 (2012). DOI: 10.1105/tpc.112.097949
Miura, K. et al. Expression profiling-based identification of CO2-responsive genes regulated by CCM1 controlling a carbon-concentrating mechanism in Chlamydomonas reinhardtii. Plant Physiol. 135, 1595–1607 (2004). DOI: 10.1104/pp.104.041400
Redekop, P. et al. Transcriptional regulation of photoprotection in dark-to-light transition- more than just a matter of excess light energy. Sci. Adv. 8, eabn1832 (2022). DOI: 10.1126/sciadv.abn1832
Colin, M. et al. Mutations affecting the mitochondrial genes encoding the cytochrome oxidase subunit I and apocytochrome b of Chlamydomonas reinhardtii. Mol. Gen. Genet. 249, 179–184 (1995). DOI: 10.1007/BF00290364
Hanawa, Y., Watanabe, M., Karatsu, Y., Fukuzawa, H. & Shiraiwa, Y. Induction of a high-CO2-inducible, periplasmic protein, H43, and its application as a high-CO2-responsive marker for study of the high-CO2-sensing mechanism in Chlamydomonas reinhardtii. Plant Cell Physiol. 48, 299–309 (2007). DOI: 10.1093/pcp/pcl066
Soupene, E. et al. Rhesus expression in a green alga is regulated by CO(2). Proc. Natl Acad. Sci. USA. 99, 7769–7773 (2002). DOI: 10.1073/pnas.112225599
Scholz, M. et al. Light-dependent N-terminal phosphorylation of LHCSR3 and LHCB4 are interlinked in Chlamydomonas reinhardtii. Plant J. 99, 877–894 (2019). DOI: 10.1111/tpj.14368
Wang, Y., Stessman, D. J. & Spalding, M. H. The CO2 concentrating mechanism and photosynthetic carbon assimilation in limiting CO2: how Chlamydomonas works against the gradient. Plant J. 82, 429–448 (2015). DOI: 10.1111/tpj.12829
Mukherjee, A. et al. Thylakoid localized bestrophin-like proteins are essential for the CO2 concentrating mechanism of Chlamydomonas reinhardtii. Proc. Natl Acad. Sci. USA. 116, 16915–16920 (2019). DOI: 10.1073/pnas.1909706116
Yoshioka, S. et al. The novel Myb transcription factor LCR1 regulates the CO2-responsive gene Cah1, encoding a periplasmic carbonic anhydrase in Chlamydomonas reinhardtii. Plant Cell 16, 1466–1477 (2004). DOI: 10.1105/tpc.021162
Aihara, Y., Fujimura-Kamada, K., Yamasaki, T. & Minagawa, J. Algal photoprotection is regulated by the E3 ligase CUL4-DDB1DET1. Nat. Plants 5, 34–40 (2019). DOI: 10.1038/s41477-018-0332-5
Trebst, A. Inhibitors in the functional dissection of the photosynthetic electron transport system. Photosyn. Res. 92, 217–224 (2007). DOI: 10.1007/s11120-007-9213-x
Maruyama, S., Tokutsu, R. & Minagawa, J. Transcriptional regulation of the stress-responsive light harvesting complex genes in Chlamydomonas reinhardtii. Plant Cell Physiol. 55, 1304–1310 (2014). DOI: 10.1093/pcp/pcu068
Eriksson, M., Villand, P., Gardeström, P. & Samuelsson, G. Induction and Regulation of Expression of a Low-CO2-Induced Mitochondrial Carbonic Anhydrase inChlamydomonas reinhardtii. Plant Physiol. 116, 637–641 (1998). DOI: 10.1104/pp.116.2.637
Roach, T., Na, C. S., Stöggl, W. & Krieger-Liszkay, A. The non-photochemical quenching protein LHCSR3 prevents oxygen-dependent photoinhibition in Chlamydomonas reinhardtii. J. Exp. Bot. 71, 2650–2660 (2020). DOI: 10.1093/jxb/eraa022
Spalding, M. H. in The molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas (eds. Rochaix, J. D., Goldschmidt-Clermont, M. & Merchant, S. S.) (1998).
Hemschemeier, A. et al. COPPER RESPONSE REGULATOR1-dependent and -independent responses of the Chlamydomonas reinhardtii transcriptome to dark anoxia. Plant Cell 25, 3186–3211 (2013). DOI: 10.1105/tpc.113.115741
Santhanagopalan, I., Wong, R., Mathur, T. & Griffiths, H. Orchestral manoeuvres in the light: crosstalk needed for regulation of the Chlamydomonas carbon concentration mechanism. J. Exp. Bot. 72, 4604–4624 (2021). DOI: 10.1093/jxb/erab169
Linthwaite, V. L. et al. The identification of carbon dioxide mediated protein post-translational modifications. Nat. Commun. 9, 3092 (2018). DOI: 10.1038/s41467-018-05475-z
Merchant, S. S. et al. The Chlamydomonas genome reveals the evolution of key animal and plant functions. Science 318, 245–250 (2007). DOI: 10.1126/science.1143609
Hasegawa, E., Hayashi, H., Asakura, S. & Kamiya, R. Stimulation of in vitro motility of Chlamydomonas axonemes by inhibition of cAMP-dependent phosphorylation. Cell Motil. Cytoskel. 8, 302–311 (1987). DOI: 10.1002/cm.970080403
Pasquale, S. M. & Goodenough, U. W. Cyclic AMP functions as a primary sexual signal in gametes of Chlamydomonas reinhardtii. J. Cell Biol. 105, 2279–2292 (1987). DOI: 10.1083/jcb.105.5.2279
Gaillard, A. R., Fox, L. A., Rhea, J. M., Craige, B. & Sale, W. S. Disruption of the A-kinase anchoring domain in flagellar radial spoke protein 3 results in unregulated axonemal cAMP-dependent protein kinase activity and abnormal flagellar motility. Mol. Biol. Cell 17, 2626–2635 (2006). DOI: 10.1091/mbc.e06-02-0095
Boonyareth, M., Saranak, J., Pinthong, D., Sanvarinda, Y. & Foster, K. W. Roles of cyclic AMP in regulation of phototaxis in Chlamydomonas reinhardtii. Biologia 64, 1058–1065 (2009). DOI: 10.2478/s11756-009-0194-4
de Montaigu, A., Sanz-Luque, E., Galván, A. & Fernández, E. A soluble guanylate cyclase mediates negative signaling by ammonium on expression of nitrate reductase in Chlamydomonas. Plant Cell 22, 1532–1548 (2010). DOI: 10.1105/tpc.108.062380
Beavo, J. A. & Brunton, L. L. Cyclic nucleotide research - still expanding after half a century. Nature 3, 710–718 (2002).
Wang, L. et al. Chloroplast-mediated regulation of CO2-concentrating mechanism by Ca2+-binding protein CAS in the green alga Chlamydomonas reinhardtii. Proc. Natl Acad. Sci. USA. 113, 12586–12591 (2016). DOI: 10.1073/pnas.1606519113
Gorman, D. S. & Levine, R. P. Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardii. Proc. Natl Acad. Sci. USA. 54, 1665–1669 (1965). DOI: 10.1073/pnas.54.6.1665
Sueoka, N. Mitotic replication of deoxyribonucleic acid in Chlamydomonas reinhardii. Proc. Natl Acad. Sci. USA. 46, 83–91 (1960). DOI: 10.1073/pnas.46.1.83
Mackinder, L. C. M. et al. A repeat protein links Rubisco to form the eukaryotic carbon-concentrating organelle. Proc. Natl Acad. Sci. U. S. A. 113, 5958–5963 (2016). DOI: 10.1073/pnas.1522866113
Gibson, D. G. et al. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Methods 6, 343–345 (2009). DOI: 10.1038/nmeth.1318
Genty, B., Briantais, J.-M. & Baker, N. R. The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim. et. Biophys. Acta (BBA) 990, 87–92 (1989). DOI: 10.1016/S0304-4165(89)80016-9
Schloss, J. A. A Chlamydomonas gene encodes a G protein beta subunit-like polypeptide. Mol. Gen. Genet. 221, 443–452 (1990). DOI: 10.1007/BF00259410
Schlesselman, J. J. Data Transformation in Two-Way Analysis of Variance. J. Am. Stat. Assoc. 10.1080/01621459.1973.10482435 (2012).
