Identification of SCF Ubiquitin Ligase Substrates by Global Protein Stability Profiling

  title={Identification of SCF Ubiquitin Ligase Substrates by Global Protein Stability Profiling},
  author={Hsueh-Chi S. Yen and Stephen J. Elledge},
  pages={923 - 929}
Ubiquitin-mediated proteolysis regulates all aspects of cellular function, and defects in this process are associated with human diseases. The limited number of identified ubiquitin ligase–substrate pairs is a major bottleneck in the ubiquitin field. We established and applied genetic technologies that combine global protein stability (GPS) profiling and genetic perturbation of E3 activity to screen for substrates of the Skp1–cullin–F-box (SCF) ubiquitin ligase in mammalian cells. Among the… 

Understanding Cullin-RING E3 Biology through Proteomics-based Substrate Identification*

Proteomic technologies for the identification of ubiquitin ligase targets are described, with a particular focus on members of the cullin-RING E3 class of ubiqu itin ligases, which use F-box proteins as substrate specific adaptor proteins.

Proteomics-based Cullin-RING E3 substrate identification

Proteomic technologies for identification of ubiquitin ligase targets with a particular focus on members of the cullin-RING E3 class of ubiqu itin ligases, which use F-box proteins as substrate specific adaptor proteins are described.

Systematic approaches to identify E3 ligase substrates

Systematic approaches to identify and validate UPS targets are highlighted and how they are underpinning rapid advances in the understanding of the biochemistry and biology of the UPS are discussed.

Enzyme–substrate relationships in the ubiquitin system: approaches for identifying substrates of ubiquitin ligases

The range of tools and techniques that can be used for substrate profiling of E3 ligases are discussed, including two-hybrid screens and co-immunoprecipitations paired with mass spectrometry.

Mechanisms and function of substrate recruitment by F-box proteins

The evolution of substrate recruitment by F-box proteins, the dysregulation of substrates in disease and potential avenues for F- box protein-directed disease therapies are focused on.

Identifying the ubiquitination targets of E6AP by orthogonal ubiquitin transfer

An orthogonal UB transfer cascade is constructed to identify specific substrates of the E3 enzyme E6AP and establishes OUT as an efficient platform to profile E3 substrates and reveal the cellular circuits mediated by the E2-E3 enzymes.

SCF Complex E3 Ubiquitin Ligases as Anticancer Targets

Recent advances in validation of SCF E3 ubiquitin ligase complex as an attractive anti-cancer target are reviewed and how MLN4924, a small molecule inhibitor of NEDD8-activating enzyme, can be developed as a novel class of anticancer agents by inhibiting SCF Ligase complex via removal of cullin neddylation is discussed.

Substrate Trapping Proteomics Reveals Targets of the βTrCP2/FBXW11 Ubiquitin Ligase

The continued use of substrate trapping proteomics to comprehensively define targets for E3 ubiquitin ligases is supported, and new functions for RAPGEF2 that may contribute to aneuploidy in cancer are established.



Ubiquitin-dependent degradation of multiple F-box proteins by an autocatalytic mechanism.

  • J. GalanM. Peter
  • Biology, Chemistry
    Proceedings of the National Academy of Sciences of the United States of America
  • 1999
The results suggest that ubiquitin-dependent degradation of F-box proteins allows rapid switching among multiple SCF complexes, thereby enabling cells to adapt quickly to changing physiological conditions and progression through different phases of the cell cycle.

A proteomic screen reveals SCFGrr1 targets that regulate the glycolytic–gluconeogenic switch

A role for SCFGrr1 in regulating the glycolytic–gluconeogenic switch is demonstrated and a proteomic screening method that uses high-throughput quantitative microscopy to comprehensively screen for ubiquitin-ligase substrates is developed.

The SCF ubiquitin ligase: insights into a molecular machine

A unifying and structurally detailed view of SCF-mediated proteolytic control of cellular processes that has been revealed by recent studies is explored.

Ubiquitin ligases: cell-cycle control and cancer

A better understanding of the ubiquitylation machinery will provide new insights into the regulatory biology of cell-cycle transitions and the development of anti-cancer drugs.

Phosphorylation-Dependent Ubiquitination of Cyclin E by the SCFFbw7 Ubiquitin Ligase

The ubiquitin ligase responsible for cyclin E ubiquitination as SCFFbw7 is identified and it is demonstrated that it is functionally conserved in yeast, flies, and mammals.

The cell-cycle regulatory protein Cks1 is required for SCFSkp2-mediated ubiquitinylation of p27

It is shown that the missing factor is CDK subunit 1 (Cks1), which belongs to the highly conserved Suc1/Cks family of proteins that bind to some CDKs and phosphorylated proteins and are essential for cell-cycle progression.