Projects per year
Ubiquitylation enzymes are involved in all aspects of eukaryotic biology and are frequently disrupted in disease. One example is the E3 ubiquitin ligase RNF12/RLIM, which is mutated in the developmental disorder Tønne-Kalscheuer syndrome (TOKAS). RNF12 TOKAS variants largely disrupt catalytic E3 ubiquitin ligase activity, which presents a pressing need to develop approaches to assess the impact of variants on RNF12 activity in patients. Here, we use photocrosslinking activity-based probes (photoABPs) to monitor RNF12 RING E3 ubiquitin ligase activity in normal and pathogenic contexts. We demonstrate that photoABPs undergo UV-induced labelling of RNF12 that is consistent with its RING E3 ligase activity. Furthermore, photoABPs robustly report the impact of RNF12 TOKAS variants on E3 activity, including variants within the RING domain and distal non-RING regulatory elements. Finally, we show that this technology can be rapidly deployed in human pluripotent stem cells. In summary, photoABPs are versatile tools that can directly identify disruptions to RING E3 ubiquitin ligase activity in human disease, thereby providing new insight into pathogenic mechanisms.
- Chemical probe
- Developmental disorder
- E3 ubiquitin ligase
- Stem cells
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology (miscellaneous)
- Health, Toxicology and Mutagenesis
- Plant Science
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- 4 Finished
RNF12 E3 Ubiquitin Ligase Substrate Degraders as a Therapeutic Strategy in Tonne-Kalscheuer Syndrome (TOKAS)
Bustos, F. & Findlay, G.
1/03/20 → 28/02/22
1/09/18 → 31/08/23
1/07/17 → 30/06/21
Student thesis: Doctoral Thesis › Doctor of Philosophy