Abstract
A significant hurdle to understanding the functions of deubiquitinases (DUBs) is the identification of their in vivo substrates. Substrate identification can be difficult for two reasons. First, many proteins that are degraded by the ubiquitin–proteasome system are expressed at relatively low levels in the cell, and second, redundancy between DUBs complicates loss of function screening approaches. Here, we describe a systematic overexpression approach that takes advantage of genome-wide resources available in S. cerevisiae to overcome these challenges and identify DUB substrates in cells.
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Acknowledgments
We thank members of the Benanti Lab for helpful discussions and suggestions on this chapter. This work was supported by National Institutes of Health grant R35GM136280 to J.A.B.
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Arsenault, H.E., Benanti, J.A. (2023). Identification of Deubiquitinase Substrates in Saccharomyces cerevisiae by Systematic Overexpression. 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_14
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DOI: https://doi.org/10.1007/978-1-0716-2803-4_14
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