Periodic Reporting for period 2 - LIP-ATG (The missing link: how do membrane lipids interplay with ATG proteins to instruct plant autophagy)
Reporting period: 2021-08-01 to 2023-01-31
We purified autophagy structures and we are now establishing their molecular profile. We already characterized their glycerophospholipid and sterol composition and further analyses are ongoing to provide an exhaustive map of the phagophore lipid composition, compare it to that of other cell compartments and identify lipids enriched in the phagophore as candidates for further functional analyses. Additionally, we established the proteome of autophagy compartments thus unraveling the presence of several lipid-related proteins including lipid-remodeling enzymes which characterization (localization, function) is in progress with the aim to explore their contribution in lipid dynamics during autophagosome formation.
We started to characterize the architectural dynamics of autophagosome formation in wild-type plants using Correlative Light Electron Microscopy combined to Electronic Tomography, including quantitative analyses of the phagophore morphology and contacts with additional subcellular compartments (see Gomez et al., Nature Communications, 2022 and unpublished). We further generated the transgenic lines and initial experiments to develop a novel technology in plant cells, aiming at exploring the repartition of key lipids and proteins within the phagophore membrane at nanometric scale.