Revealing ?-TrCP activity dynamics in live cells with a genetically encoded biosensor.

The F-box protein beta-transducin repeat containing protein (?-TrCP) acts as a substrate adapter for the SCF E3 ubiquitin ligase complex, plays a crucial role in cell physiology, and is often deregulated in many types of cancers. Here, we develop a fluorescent biosensor to quantitatively measure ?-TrCP activity in live, single ...
cells in real-time. We find ?-TrCP remains constitutively active throughout the cell cycle and functions to maintain discreet steady-state levels of its substrates. We find no correlation between expression levels of ?-TrCP and ?-TrCP activity, indicating post-transcriptional regulation. A high throughput screen of small-molecules using our reporter identifies receptor-tyrosine kinase signaling as a key axis for regulating ?-TrCP activity by inhibiting binding between ?-TrCP and the core SCF complex. Our study introduces a method to monitor ?-TrCP activity in live cells and identifies a key signaling network that regulates ?-TrCP activity throughout the cell cycle.
Mesh Terms:
Biosensing Techniques, F-Box Proteins, Protein-Tyrosine Kinases, Ubiquitin-Protein Ligases, beta-Transducin Repeat-Containing Proteins
Nat Commun
Date: Oct. 26, 2022
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