Abstract
Ubiquitin modification is known to regulate endocytic trafficking of many different types of cargo in eukaryotic cells, but it can be challenging to determine what role, if any, ubiquitin plays in the trafficking of a novel or uncharacterized endocytic cargo. Here, we describe a useful approach that leverages fusion to deubiquitinase (DUB) catalytic domains to explore the role ubiquitin plays in endocytic trafficking. This approach can be applied to the analysis of many different endocytic cargos in different cell types, and it can also be used to study linkage specificity in endocytic trafficking. Several different trafficking assays are described to illustrate the broad utility of this “DUB fusion” approach.
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Acknowledgments
This work was supported by NIH grant R35GM144112 (to JAM).
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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Hepowit, N.L., Denise, A.S., MacGurn, J.A. (2023). Use of Deubiquitinase Fusion Proteins to Characterize Endocytic Trafficking in Yeast. In: Maupin-Furlow, J., Edelmann, M.J. (eds) Deubiquitinases. Methods in Molecular Biology, vol 2591. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2803-4_17
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DOI: https://doi.org/10.1007/978-1-0716-2803-4_17
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