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pSILAC mass spectrometry reveals ZFP91 as IMiD-dependent substrate of the CRL4CRBN ubiquitin ligase.

  • An J Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA.
  • Ponthier CM Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA.
  • Sack R Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, CH-4058 Basel, Switzerland.
  • Seebacher J Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, CH-4058 Basel, Switzerland.
  • Stadler MB Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, CH-4058 Basel, Switzerland.
  • Donovan KA Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA.
  • Fischer ES Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02215, USA.
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  • 2017-05-23
Published in:
  • Nature communications. - 2017
Amino Acid Motifs
Animals
Antineoplastic Agents
Gene Deletion
HCT116 Cells
HEK293 Cells
Humans
Lenalidomide
Mass Spectrometry
Mice
Multiple Myeloma
Myelodysplastic Syndromes
Peptide Hydrolases
Proteomics
Substrate Specificity
Ubiquitin
Ubiquitin-Protein Ligases
Ubiquitination
Zinc Fingers
English Thalidomide and its derivatives lenalidomide and pomalidomide (IMiDs) are effective treatments of haematologic malignancies. It was shown that IMiDs impart gain-of-function properties to the CUL4-RBX1-DDB1-CRBN (CRL4CRBN) ubiquitin ligase that enable binding, ubiquitination and degradation of key therapeutic targets such as IKZF1, IKZF3 and CSNK1A1. While these substrates have been implicated as efficacy targets in multiple myeloma (MM) and 5q deletion associated myelodysplastic syndrome (del(5q)-MDS), other targets likely exist. Using a pulse-chase SILAC mass spectrometry-based proteomics approach, we demonstrate that lenalidomide induces the ubiquitination and degradation of ZFP91. We establish ZFP91 as a bona fide IMiD-dependent CRL4CRBN substrate and further show that ZFP91 harbours a zinc finger (ZnF) motif, related to the IKZF1/3 ZnF, critical for IMiD-dependent CRBN binding. These findings demonstrate that single time point pulse-chase SILAC mass spectrometry-based proteomics (pSILAC MS) is a sensitive approach for target identification of small molecules inducing selective protein degradation.
Language
  • English
Open access status
gold
Identifiers
Persistent URL
https://folia.unifr.ch/global/documents/102325
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