Project description
Dissecting the mechanism of autophagy
Autophagy refers to the self-eating or self-digestion of cellular components and is a highly complex and dynamic process that contributes to cellular and organismal homeostasis, adaptation to stress, and maintenance of cellular health. Damaged or unnecessary components are sequestered within specialised double-membrane structures called autophagosomes, which then fuse with lysosomes. Understanding autophagy is crucial for protecting organisms from diseases like neurodegeneration, cancer, and infections. Funded by the European Research Council, the AutoRecon project aims to investigate the mechanisms of autophagosome formation in eukaryotic cells using biochemical and cell biological techniques. Researchers will determine key factors and their mode of action, providing unprecedented insight into de novo organelle formation.
Objective
I propose to study how eukaryotic cells generate autophagosomes, organelles bounded by a double membrane. These are formed during autophagy and mediate the degradation of cytoplasmic substances within the lysosomal compartment. Autophagy thereby protects the organism from pathological conditions such as neurodegeneration, cancer and infections. Many core factors required for autophagosome formation have been identified but the order in which they act and their mode of action is still unclear. We will use a combination of biochemical and cell biological approaches to elucidate the choreography and mechanism of these core factors. In particular, we will focus on selective autophagy and determine how the autophagic machinery generates an autophagosome that selectively contains the cargo.
To this end we will focus on the cytoplasm-to-vacuole-targeting pathway in S. cerevisiae that mediates the constitutive delivery of the prApe1 enzyme into the vacuole. We will use cargo mimetics or prApe1 complexes in combination with purified autophagy proteins and vesicles to reconstitute the process and so determine which factors are both necessary and sufficient for autophagosome formation, as well as elucidating their mechanism of action.
In parallel we will study selective autophagosome formation in human cells. This will reveal common principles and special adaptations. In particular, we will use cell lysates from genome-edited cells in combination with purified autophagy proteins to reconstitute selective autophagosome formation around ubiquitin-positive cargo material. The insights and hypotheses obtained from these reconstituted systems will be validated using cell biological approaches.
Taken together, our experiments will allow us to delineate the major steps of autophagosome formation during selective autophagy. Our results will yield detailed insights into how cells form and shape organelles in a de novo manner, which is major question in cell- and developmental biology.
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Programme(s)
Funding Scheme
ERC-COG - Consolidator GrantHost institution
1010 Wien
Austria