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Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor
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-T...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9606124/ https://www.ncbi.nlm.nih.gov/pubmed/36289220 http://dx.doi.org/10.1038/s41467-022-33762-3 |
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author | Paul, Debasish Kales, Stephen C. Cornwell, James A. Afifi, Marwa M. Rai, Ganesha Zakharov, Alexey Simeonov, Anton Cappell, Steven D. |
author_facet | Paul, Debasish Kales, Stephen C. Cornwell, James A. Afifi, Marwa M. Rai, Ganesha Zakharov, Alexey Simeonov, Anton Cappell, Steven D. |
author_sort | Paul, Debasish |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9606124 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-96061242022-10-28 Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor Paul, Debasish Kales, Stephen C. Cornwell, James A. Afifi, Marwa M. Rai, Ganesha Zakharov, Alexey Simeonov, Anton Cappell, Steven D. Nat Commun Article 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. Nature Publishing Group UK 2022-10-26 /pmc/articles/PMC9606124/ /pubmed/36289220 http://dx.doi.org/10.1038/s41467-022-33762-3 Text en © This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Paul, Debasish Kales, Stephen C. Cornwell, James A. Afifi, Marwa M. Rai, Ganesha Zakharov, Alexey Simeonov, Anton Cappell, Steven D. Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor |
title | Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor |
title_full | Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor |
title_fullStr | Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor |
title_full_unstemmed | Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor |
title_short | Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor |
title_sort | revealing β-trcp activity dynamics in live cells with a genetically encoded biosensor |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9606124/ https://www.ncbi.nlm.nih.gov/pubmed/36289220 http://dx.doi.org/10.1038/s41467-022-33762-3 |
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