Abstract
Budding yeast Saccharomyces cerevisiae is an ideal model organism to study membrane trafficking pathways. The ESCRT (endosomal sorting complexes required for transport) pathway was first identified in this organism. Upon recognition of endocytosed ubiquitinated membrane proteins at endosomes, ESCRTs assemble at these organelles to catalyze the biogenesis of multivesicular bodies (MVBs). Formation of MVBs leads to the trafficking of these membrane proteins to vacuoles for degradation. Here, we describe genetic and biochemical approaches to study ESCRT function. We outline in vivo endocytosis assays using two model cargoes in Saccharomyces cerevisiae and also describe an in vitro approach to analyze ESCRT-III polymerization on lipid monolayers.
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Acknowledgments
Work in the Emr lab is supported by a Cornell University Research Grant CU3704. Sudeep Banjade is a HHMI fellow of the Damon Runyon Cancer Research Foundation (DRG-2273-16). Shaogeng Tang is a Merck fellow of the Damon Runyon Cancer Research Foundation (DRG-2301-17). We thank all members of the Emr lab for building these protocols in the lab over the years.
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Banjade, S., Tang, S., Emr, S.D. (2019). Genetic and Biochemical Analyses of Yeast ESCRT. In: Culetto, E., Legouis, R. (eds) The ESCRT Complexes. Methods in Molecular Biology, vol 1998. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9492-2_8
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DOI: https://doi.org/10.1007/978-1-4939-9492-2_8
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