|Morphogenesis, Signaling, Modeling|
|Dynamics and Expression of plant Genomes|
|Adaptation of Plants to the Environment|
|Reproduction and Seeds|
|Plant cell wall, function and utilization|
Dynamics and structure of
Keywords :Lipid bodies, oleosome, dynamic, function, proteomic, crystallography, biophysics, low soluble proteins, protein interaction, lipid design, Arabidopsis thaliana, Brassica napus, Saccharomyces cerevisiae, Yarrowia lipolytica, Streptomyces.
|Doctoral school affiliation : ED 435 ABIES|
Germination and seedling establishment require high energy input and the supply of carbon backbones for various biosynthetic pathways such as that of lipids. In effect, the plant organ exhibiting the highest levels of lipid accumulation is seeds. Storage lipids are accumulated in lipid bodies (LBs), also called lipid droplets or oil bodies. LBs structure is conserved among living organisms, with a core composed of neutral lipids surrounded by a monolayer of phospholipids, in which a highly variable number of proteins is embedded. These proteins in microbe or animal LBs are mostly comprised of enzymes, or proteins involved in subcellular transport. Seed LBs contain a limited set of proteins, with few enzymes. In particular, little is known about the dynamics of LBs during seed life cycle. To date, no structure has been determined to high resolution for a protein inserted in a phospholipid monolayer. To identify and characterize proteins contributing to the dynamic and the structure of LBs, we apply genetic, biochemical, molecular, and structural approaches. As models we use oleaginous plants (Arabidopsis thaliana, Brassica napus), and also yeasts (Saccharomyces cerevisae, Yarrowia lipolytica).
main goals are:
accordance with the principles of “Green Chemistry”, our research
aims to modulate amounts and tailor the nature of lipids produced, as
well as to facilitate storage lipid extraction.
Main Results :
Our efforts are focused on two investigative
axes: « Structural and functional », and « Proteomic
and dynamic ». Plants and microorganisms able to store lipids are
used as models for our studies.
have expertise in the determination of protein post-translational modifications.
We proved that A. thaliana caleosin was phosphorylated (Purkrtova
2007), and that its interfacial properties were modified upon calcium
binding (Purkrtova 2008). Using structural proteomic approaches, the topology
of proteins at the surface of lipid bodies has also been determined. (Vermachova).
Compared to microbial or animal lipid bodies, (Athenstaedt 2006), LBs from A. thaliana or B. napus mature seed seem to be in a quiescent state (Jolivet 2004, 2009, d’Andréa 2007b, Popluechai submitted). Rapeseed LB proteins follow a sequential expression pattern, so it is likely that LBs undergo an ordered maturation scheme (Jolivet, unpublished).
Significant collaborations, contracts
Main collaborations: UMR INRA-INSA LISBP Toulouse, ESPCI ParisTech, ENSCP Paris, Institut Pasteur, Synchrotron Soleil, UMR AgroParisTech-INRA Micalis Grignon, UMR AgroCampus INRA APBV Rennes, UR INRA BIA Nantes, University Paris XI, Graz, Prague, Olomouc, Pittsburg; proteomic facility PAPPSO Le Moulon, CETIOM Pessac.
Selected Publications :
Baud A, Ayme L, Gonnet F, Salard I, Gohon Y, Jolivet P, Brodolin K, Da Silva P, Giuliani A, Sclavi B, Chardot T, Mercere P, Roblin P, Daniel R (2017) SOLEIL shining on the solution-state structure of biomacromolecules by synchrotron X-ray footprinting at the Metrology beamline. Journal of Synchrotron Radiation 24: 576-585
Groeme R., Airouche S., Kope?ný D., Jaekel J., Savko M., Berjont N., Bussieres L., Le Mignon M., Jagic F., Zieglmayer P., Baron-Bodo V., Bordas-Le Floch V., Mascarell L., Briozzo P., Moingeon P. (2016) Structural and Functional Characterization of the Major Allergen Amb a 11 from Short Ragweed Pollen. J. Biol. Chem. 291 : 13076-13087Haili N, Louap J, Canonge M, Jagic F, Louis-Mondesir C, Chardot T, Briozzo P (2016) Expression of Soluble Forms of Yeast Diacylglycerol Acyltransferase 2 That Integrate a Broad Range of Saturated Fatty Acids in Triacylglycerols. PLoS One 11: e0165431
Kopecný D., Koncitíková R., Popelká H., Briozzo P., Vigouroux A., Kopecná M, Zalabák D., Šebela M., Skopalová J., Frébort I., Moréra S. (2016) Kinetic and structural investigation of the cytokinin oxidase/dehydrogenase active site. FEBS J. 283: 361-377
Deruyffelaere C, Bouchez I, Morin H, Guillot A, Miquel M, Froissard M, Chardot T, D'Andrea S (2015) Ubiquitin-Mediated Proteasomal Degradation of Oleosins is Involved in Oil Body Mobilization During Post-Germinative Seedling Growth in Arabidopsis. Plant Cell Physiol. 56: 1374-1387
Boulard C, Bardet M, Chardot T, Dubreucq B, Gromova M, Guillermo A, Miquel M, Nesi N, Yen-Nicolay S, Jolivet P (2015) The structural organization of seed oil bodies could explain the contrasted oil extractability observed in two rapeseed genotypes. Planta 242: 53-68
Bouchez I, Pouteaux M, Canonge M, Genet M, Chardot T, Guillot A, Froissard M (2015) The Rab7-like protein Ypt7p is involved in Saccharomyces cerevisiae lipid droplet dynamics. Biolopen 4: 764-775
Ayme L, Nicaud J-M, Jolivet P, Chardot T (2015) Molecular characterization of the Elaeis guineensis medium-chain fatty acid diacylglycerol acyltransferase DGAT1-1 by heterologous expression in Yarrowia lipolytica. PLoS One 10: e0143113 (pdf) Communiqué de presse INRA
Vermachova M, Purkrtova Z, Jolivet P, Chardot T, Kodicek M (2014) Combining chymotrypsin/trypsin digestion to identify hydrophobic proteins from oil bodies. In Plant Proteomics. Methods and Protocols. Se49
Ayme L, Baud S, Dubreucq B, Joffre F, Chardot T (2014) Function and Localization of the Arabidopsis thaliana Diacylglycerol Acyltransferase DGAT2 Expressed in Yeast. PLoS One 9: e92237 (pdf) Communiqué de press INRA
Le Marechal P, Decottignies P, Marchand CH, Degrouard J, Jaillard D, Dulermo T, Froissard M, Smirnov A, Chapuis V, Virolle MJ (2013) Comparative Proteomic Analysis of Streptomyces lividans Wild-Type and ppk Mutant Strains Reveals the Importance of Storage Lipids for Antibiotic Biosynthesis. Appl Environ Microbiol 79: 5907-5917 (pdf)
Haïli N, Arnal N, Quadrado M, Amiar S, Tcherkez G, Dahan J, Briozzo P, Colas des Francs Small C, Vrielynck N, Mireau H (2013) The pentatricopeptide repeat MTSF1 protein stabilizes the nad4 mRNA in Arabidopsis mitochondria. Nucleic Acids Res 41:6650-6663 (pdf)
Jamme F, Vindigni JD, Méchin V, Cherifi T, Chardot T, Froissard, M. (2013) Single cell synchrotron FT-IR microspectroscopy reveals a link between neutral lipid and storage carbohydrate fluxes in S. cerevisiae. PLOS ONE. DOI: 10.1371/journal.pone.0074421 (pdf) Actualité Centre INRA Versailles-Grignon
© INRA 2010