Functional E3 ligase hotspots and resistance mechanisms to small-molecule degraders

DOI: 10.1038/s41589-022-01177-2

Authors: Alexander Hanzl (CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences) , Ryan Casement (University of Dundee) , Hana Imrichova (CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences) , Scott J. Hughes (University of Dundee) , Eleonora Barone (CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences) , Andrea Testa (University of Dundee) , Sophie Bauer (CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences) , Jane Wright (University of Dundee) , Matthias Brand (University of Dundee) , Alessio Ciulli (University of Dundee) , Georg E. Winter (CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Chemical Biology , VOL 580

State: Published (Approved)
Published: November 2022
Diamond Proposal Number(s): 14980

Abstract: Targeted protein degradation is a novel pharmacology established by drugs that recruit target proteins to E3 ubiquitin ligases. Based on the structure of the degrader and the target, different E3 interfaces are critically involved, thus forming defined ‘functional hotspots’. Understanding disruptive mutations in functional hotspots informs on the architecture of the assembly, and highlights residues susceptible to acquire resistance phenotypes. Here we employ haploid genetics to show that hotspot mutations cluster in substrate receptors of hijacked ligases, where mutation type and frequency correlate with gene essentiality. Intersection with deep mutational scanning revealed hotspots that are conserved or specific for chemically distinct degraders and targets. Biophysical and structural validation suggests that hotspot mutations frequently converge on altered ternary complex assembly. Moreover, we validated hotspots mutated in patients that relapse from degrader treatment. In sum, we present a fast and widely accessible methodology to characterize small-molecule degraders and associated resistance mechanisms.

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I24-Microfocus Macromolecular Crystallography

Added On: 09/11/2022 08:57

Discipline Tags:

Non-Communicable Diseases Health & Wellbeing Cancer Biochemistry Genetics Chemistry Structural biology Biophysics Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)