Descripción del proyecto
Análisis del mecanismo de la autofagia en plantas
La autofagia se refiere a al proceso de autoalimentación o de autodigestión de componentes celulares y es un mecanismo altamente complejo y dinámico, que contribuye a la homeostasis celular y del organismo, a la adaptación al estrés y al mantenimiento de la salud celular. Los componentes dañados o innecesarios se secuestran dentro de unas estructuras especializadas de doble membrana denominadas autofagosomas, que luego se fusionan con los lisosomas. Comprender la autofagia es crucial para proteger a los organismos de enfermedades como la neurodegeneración, las neoplasias y las infecciones. El equipo del proyecto AutoRecon, financiado con fondos del Consejo Europeo de Investigación (ERC, por sus siglas en inglés), pretende investigar los mecanismos de formación de autofagosomas en células eucariotas mediante técnicas bioquímicas y de biología celular. Los investigadores definirán los factores clave y su modo de acción, lo que proporcionará una perspectiva sin precedentes de la formación de novo de orgánulos.
Objetivo
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.
Ámbito científico
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ERC-COG - Consolidator GrantInstitución de acogida
1010 Wien
Austria