TY - JOUR T1 - Systematic multi-level analysis of an organelle proteome reveals new peroxisomal functions JF - bioRxiv DO - 10.1101/2021.12.08.471723 SP - 2021.12.08.471723 AU - Eden Yifrach AU - Duncan Holbrook-Smith AU - Jérôme Bürgi AU - Alaa Othman AU - Miriam Eisenstein AU - Carlo W.T Van Roermund AU - Wouter Visser AU - Asa Tirosh AU - Chen Bibi AU - Shahar Galor AU - Uri Weil AU - Amir Fadel AU - Yoav Peleg AU - Hans R Waterham AU - Ronald J A Wanders AU - Matthias Wilmanns AU - Nicola Zamboni AU - Maya Schuldiner AU - Einat Zalckvar Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/12/11/2021.12.08.471723.abstract N2 - Seventy years following the discovery of peroxisomes, their proteome remains undefined. Uncovering the complete peroxisomal proteome, the peroxi-ome, is crucial for understanding peroxisomal activities and cellular metabolism. We used high- content microscopy to uncover the peroxi-ome of the model eukaryote – Saccharomyces cerevisiae. This strategy enabled us to expand the known organellar proteome by ∼40% and paved the way for performing systematic, whole-organellar proteome assays. Coupled with targeted experiments this allowed us to discover new peroxisomal functions. By characterizing the sub-organellar localization and protein targeting dependencies into the organelle, we unveiled non-canonical targeting routes. Metabolomic analysis of the peroxi-ome revealed the role of several newly-identified resident enzymes. Importantly, we found a regulatory role of peroxisomes during gluconeogenesis, which is fundamental for understanding cellular metabolism. With the current recognition that peroxisomes play a crucial part in organismal physiology, our approach lays the foundation for deep characterization of peroxisome function in health and disease. ER -