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

Author

Listed:
  • Debasish Paul

    (National Cancer Institute)

  • Stephen C. Kales

    (National Institutes of Health)

  • James A. Cornwell

    (National Cancer Institute)

  • Marwa M. Afifi

    (National Cancer Institute)

  • Ganesha Rai

    (National Institutes of Health)

  • Alexey Zakharov

    (National Institutes of Health)

  • Anton Simeonov

    (National Institutes of Health)

  • Steven D. Cappell

    (National Cancer Institute)

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.

Suggested Citation

  • Debasish Paul & Stephen C. Kales & James A. Cornwell & Marwa M. Afifi & Ganesha Rai & Alexey Zakharov & Anton Simeonov & Steven D. Cappell, 2022. "Revealing β-TrCP activity dynamics in live cells with a genetically encoded biosensor," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-33762-3
    DOI: 10.1038/s41467-022-33762-3
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    Cited by:

    1. Linda Lauinger & Anna Andronicos & Karin Flick & Clinton Yu & Geetha Durairaj & Lan Huang & Peter Kaiser, 2024. "Cadmium binding by the F-box domain induces p97-mediated SCF complex disassembly to activate stress response programs," Nature Communications, Nature, vol. 15(1), pages 1-13, December.

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