Details

Autor(en) / Beteiligte

• Geffen, Yifat
• Appleboim, Alon
• Gardner, Richard G.
• Friedman, Nir
• Sadeh, Ronen
• Ravid, Tommer

Titel

Mapping the Landscape of a Eukaryotic Degronome

Ist Teil von

• Molecular cell, 2016-09, Vol.63 (6), p.1055-1065

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2016

Quelle

Access via ScienceDirect (Elsevier)

Beschreibungen/Notizen

• 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. [Display omitted] •New reporter-based high-throughput degradation assay was developed•Thousands of degradation signals (degrons) from the yeast transcriptome were identified•A secondary screen revealed multiple targets of the E3 ligase Doa10p•The addition of nuclear localization signal greatly changed degron propensity Using a reporter-based competition assay, Geffen, Appleboim et al. measured the degradation potency of thousands of polypeptides. The assay was further used for identifying new quality control substrates of the ubiquitin system and for comparing cytosolic and nuclear degradation potency.