Nuclear mTORC1 Live-Cell Sensor nTORSEL Reports Differential Nuclear mTORC1 Activity in Cell Lines
Yifan Wang1, Canrong Li1, Yingyi Ouyang1
1School of Medicine, Shenzhen Campus of Sun Yat-Sen University, Sun Yat-Sen University, Shenzhen 518107, China.
International Journal of Molecular Sciences
|November 27, 2024
Summary
Researchers developed nTORSEL, a novel sensor to monitor nuclear mechanistic target of rapamycin complex 1 (mTORC1) activity. This tool aids in understanding nuclear mTORC1 signaling pathways and their biological functions.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Mammalian/mechanistic target of rapamycin complex 1 (mTORC1) regulates cellular processes.
- mTORC1 activation occurs at lysosomes, phosphorylating substrates in the nucleus, cytosol, and lysosomes.
- Nuclear mTORC1 (nmTORC1) signaling and functions remain poorly understood due to limited monitoring tools.
Purpose of the Study:
- To develop a genetically encoded sensor for monitoring nmTORC1 activity.
- To investigate the signaling and biological functions of nmTORC1.
- To provide a tool for studying nmTORC1 in various cellular contexts.
Main Methods:
- Development of a genetically encoded nmTORC1 sensor (nTORSEL) based on eIF4E binding protein 1 (4EBP1) phosphorylation.
- Utilizing fluorescent microscopy and biochemical analyses for validation.
- Testing nTORSEL in cell lines with varying nmTORC1 activity levels.
Main Results:
- nTORSEL successfully detects nmTORC1 activity by monitoring 4EBP1 phosphorylation in the nucleus.
- The sensor exhibits a punctate nuclear pattern dependent on nmTORC1 activity.
- nTORSEL specifically identified inhibition of nmTORC1 in mouse embryonic fibroblast (MEF) cells.
Conclusions:
- nTORSEL is an effective tool for investigating nmTORC1 signaling.
- The sensor enables specific detection of nmTORC1 activity and inhibition.
- This tool facilitates further research into the roles of nmTORC1 in cellular functions.
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