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

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

Authors :
Paul D
Kales SC
Cornwell JA
Afifi MM
Rai G
Zakharov A
Simeonov A
Cappell SD
Source :
Nature communications [Nat Commun] 2022 Oct 26; Vol. 13 (1), pp. 6364. Date of Electronic Publication: 2022 Oct 26.
Publication Year :
2022

Abstract

The F-box protein beta-transducin repeat containing protein (β-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 β-TrCP activity in live, single cells in real-time. We find β-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 β-TrCP and β-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 β-TrCP activity by inhibiting binding between β-TrCP and the core SCF complex. Our study introduces a method to monitor β-TrCP activity in live cells and identifies a key signaling network that regulates β-TrCP activity throughout the cell cycle.<br /> (© 2022. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Details

Language :
English
ISSN :
2041-1723
Volume :
13
Issue :
1
Database :
MEDLINE
Journal :
Nature communications
Publication Type :
Academic Journal
Accession number :
36289220
Full Text :
https://doi.org/10.1038/s41467-022-33762-3