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Updated: Sep 9, 2025
14:57
Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
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Pharmacologic interrogation of USP28 cellular function in p53 signaling
Ariana S Bratt1, Susan Kilgas2, Maria I Tarazona Guzman1
1Department of Cancer Biology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Biological Chemistry & Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
Cell Chemical Biology
|September 3, 2025
Summary
Researchers developed CAS-010, a specific inhibitor for USP28 (a deubiquitinating enzyme), to study its role in p53 signaling and cell fate. This chemical probe aids in understanding ubiquitin signaling pathways.
Area of Science:
- Biochemistry
- Molecular Biology
- Chemical Biology
Background:
- Deubiquitinating enzymes (DUBs) regulate critical cellular processes including protein degradation.
- Understanding DUB function is hindered by a lack of specific chemical probes.
- USP28 is a DUB implicated in various cellular pathways, but its precise roles require further investigation.
Purpose of the Study:
- To develop and validate a specific chemical probe for USP28.
- To investigate the role of USP28 in p53 transactivation and the 53BP1-USP28 interaction.
- To provide tools for studying USP28's function in cell cycle control and cell fate.
Main Methods:
- Development of CAS-010, a ubiquitin-competitive USP28 inhibitor.
- Structure-based drug design and SAR analysis for inhibitor selectivity.
- Biochemical assays to determine inhibitor potency and selectivity.
- Cell-based assays to validate on-target effects on p53 transactivation.
- Investigation of the impact of CAS-010 on the 53BP1-USP28 interaction.
Main Results:
- CAS-010 identified as a low nanomolar, selective inhibitor of USP28 with some activity against USP25.
- Crystal structure rationalized inhibitor selectivity and SAR trends.
- Validated on-target inhibition of p53 transactivation by CAS-010.
- Demonstrated that CAS-010 disrupts the 53BP1-USP28 interaction, highlighting USP28's catalytic role.
- Developed a negative control (WPT-086) and inhibitor-resistant mutant for further studies.
Conclusions:
- CAS-010 is a well-validated chemical tool for studying USP28.
- USP28 catalytic activity is crucial for the 53BP1-USP28 interaction.
- These tools will advance research into USP28's role in p53-mediated cell cycle control and cell fate determination.
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