AI Article Synopsis
- * The study introduces photocrosslinking activity-based probes (photoABPs) to assess RNF12's E3 ubiquitin ligase activity, showing that these probes can effectively label RNF12 and reveal how TOKAS variants alter its function.
- * The findings suggest photoABPs can be quickly applied to human pluripotent stem cells, serving as powerful tools for identifying dysfunctions in E3 ubiquitin ligase activity related to human diseases, thus enhancing our understanding of
Article Abstract
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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9240097 | PMC |
| http://dx.doi.org/10.26508/lsa.202101248 | DOI Listing |
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