Pyrenoid functions revealed by proteomics in Chlamydomonas reinhardtii

Yu Zhan , Christophe H. Marchand (1) , Alexandre Maes (1) , Adeline Mauriès (1) , Yi Sun , James S Dhaliwal , James Uniacke , Simon Arragain , Heng Jiang , Nicholas D. Gold , Vincent J.J. Martin , Stéphane D. Lemaire (1) , William Zerges

1 LBMCE - Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes

Organelles are intracellular compartments which are themselves compartmentalized. Bio-genic and metabolic processes are localized to specialized domains or microcompartments to enhance their efficiency and suppress deleterious side reactions. An example of intra-organellar compartmentalization is the pyrenoid in the chloroplasts of algae and hornworts. This microcompartment enhances the photosynthetic CO 2-fixing activity of the Calvin-Ben-son cycle enzyme Rubisco, suppresses an energetically wasteful oxygenase activity of Rubisco, and mitigates limiting CO 2 availability in aquatic environments. Hence, the pyre-noid is functionally analogous to the carboxysomes in cyanobacteria. However, a comprehensive analysis of pyrenoid functions based on its protein composition is lacking. Here we report a proteomic characterization of the pyrenoid in the green alga Chlamydomonas rein-hardtii. Pyrenoid-enriched fractions were analyzed by quantitative mass spectrometry. Contaminant proteins were identified by parallel analyses of pyrenoid-deficient mutants. This pyrenoid proteome contains 190 proteins, many of which function in processes that are known or proposed to occur in pyrenoids: e.g. the carbon concentrating mechanism, starch metabolism or RNA metabolism and translation. Using radioisotope pulse labeling experiments , we show that pyrenoid-associated ribosomes could be engaged in the localized synthesis of the large subunit of Rubisco. New pyrenoid functions are supported by proteins in tetrapyrrole and chlorophyll synthesis, carotenoid metabolism or amino acid metabolism. Hence, our results support the long-standing hypothesis that the pyrenoid is a hub for metabolism. The 81 proteins of unknown function reveal candidates for new participants in these processes. Our results provide biochemical evidence of pyrenoid functions and a resource for future research on pyrenoids and their use to enhance agricultural plant productivity. Data are available via ProteomeXchange with identifier PXD004509.

Domaines: Sciences du Vivant [q-bio]

Yu Zhan, Christophe H. Marchand, Alexandre Maes, Adeline Mauriès, Yi Sun, et al.. Pyrenoid functions revealed by proteomics in Chlamydomonas reinhardtii. PLoS ONE, 2018, 13 (2), pp.e0185039. ⟨10.1371/journal.pone.0185039⟩. ⟨hal-01744153⟩