Dissection of molecular Mechanisms regulating Pexophagy in human Cells

• Applicant: Privatdozent Dr. Andreas Till
• Subject Area: Cell Biology
• Term: from 2009 to 2012
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 146679195

Peroxisomes are single-membrane –bound organelles that contribute to several essential metabolic pathways including b-oxidation of fatty acids and detoxication processes. Disturbance in peroxisome functionality is associated with complex human diseases characterized by severe metabolic defects and premature mortality. Superfluous or dysfunctional peroxisomes are degraded by proteolytic pro-cesses that use molecular components of the autophagy machinery. This selective degradation of peroxisomes (´pexophagy´) has been extensively studied in yeast resulting in the identification of several yeast genes that execute or regulate pexophagy. In contrast, the molecular key players that regulate pexophagy in mammalian cells are largely unknown. This project aims to use an RNAi - based High Throughput Screening approach to identify gene networks that contribute to pexophagy regulation in human cells. After establishment of experimental procedures suitable to induce and to monitor pexophagy, a human siRNA library will be screened using automated image analysis. Confirmed candidate genes will be validated for their specific role in human pexophagy by applying different functional approaches. Finally, the yeast orthologues of selected candidates genes will be used to generate yeast knock-out strains for comparative analysis of pexophagy regulation in yeast and mammalian cells. The data derived from the screening approach followed by functional assessment will help to build up a functional road map of pexophagy regulation in different phylogenetic taxa.

• DFG Programme: Research Fellowships
• International Connection: USA
• Host: Professor Dr. Suresh Subramani