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Mapping the landscape of a eukaryotic degronome

Item Type:Article
Title:Mapping the landscape of a eukaryotic degronome
Creators Name:Geffen, Y. and Appleboim, A. and Gardner, R.G. and Friedman, N. and Sadeh, R. and Ravid, T.
Abstract:The ubiquitin-proteasome system (UPS) for protein degradation has been under intensive study, and yet, we have only partial understanding of mechanisms by which proteins are selected to be targeted for proteolysis. One of the obstacles in studying these recognition pathways is the limited repertoire of known degradation signals (degrons). To better understand what determines the susceptibility of intracellular proteins to degradation by the UPS, we developed an unbiased method for large-scale identification of eukaryotic degrons. Using a reporter-based high-throughput competition assay, followed by deep sequencing, we measured a degradation potency index for thousands of native polypeptides in a single experiment. We further used this method to identify protein quality control (PQC)-specific and compartment-specific degrons. Our method provides an unprecedented insight into the yeast degronome, and it can readily be modified to study protein degradation signals and pathways in other organisms and in various settings.
Keywords:Binding Sites, Chromosome Mapping, Fungal Gene Expression Regulation, Fungal Genome, Gene Library, High-Throughput Screening Assays, Phosphorylation, Proteasome Endopeptidase Complex, Protein Binding, Protein Folding, Protein Interaction Domains and Motifs, Proteolysis, Proteome, Saccharomyces Cerevisiae, Saccharomyces Cerevisiae Proteins, Ubiquitin, Ubiquitin-Protein Ligases, alpha-Helical Protein Conformation
Source:Molecular Cell
ISSN:1097-2765
Publisher:Cell Press
Volume:63
Number:6
Page Range:1055-1065
Date:15 September 2016
Official Publication:https://doi.org/10.1016/j.molcel.2016.08.005
PubMed:View item in PubMed

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