PT  - JOURNAL ARTICLE
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
TI  - Systematic multi-level analysis of an organelle proteome reveals new peroxisomal functions
AID  - 10.1101/2021.12.08.471723
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.12.08.471723
4099  - http://biorxiv.org/content/early/2021/12/17/2021.12.08.471723.short
4100  - http://biorxiv.org/content/early/2021/12/17/2021.12.08.471723.full
AB  - 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.Competing Interest StatementThe authors have declared no competing interest.