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Light-gated integrator for highlighting kinase activity in living cells
Wei Lin1, Abhishek Phatarphekar2, Yanghao Zhong2,3
1Department of Pharmacology, University of California San Diego, La Jolla, CA, USA. wel328@health.ucsd.edu.
Nature Communications
|September 6, 2024
Summary
Researchers developed a new tool, KINACT, to permanently record dynamic kinase signals as fluorescent marks. This innovation aids in understanding disease mechanisms and developing targeted therapies by enabling robust monitoring and manipulation of kinase activity.
Area of Science:
- Molecular Biology
- Cell Signaling
- Biotechnology
Background:
- Protein kinases are crucial regulators of biological processes and disease.
- Current fluorescent biosensors allow real-time visualization but lack recording capabilities.
- New tools are needed for post hoc analysis and manipulation of transient kinase signaling.
Purpose of the Study:
- To develop a novel molecular tool for recording dynamic kinase activity.
- To enable permanent fluorescent marking of transient signaling events.
- To facilitate deeper insights into disease mechanisms and therapeutic targeting.
Main Methods:
- Development of a light-gated kinase activity coupled transcriptional integrator (KINACT).
- Utilizing KINACT to convert dynamic kinase signals into permanent fluorescent marks.
- Application of KINACT for monitoring subcellular PKA activity and 3D culture distributions.
- Employing KINACT for high-throughput screening of kinase activators and inhibitors.
Main Results:
- KINACT enables robust, scalable monitoring of kinase activity.
- Accurate recording of subcellular Protein Kinase A (PKA) activity.
- Visualization of PKA activity distribution in 3D cell cultures.
- Successful identification of PKA activators and inhibitors via high-throughput screening.
- Feedback manipulation of oncogenic G protein signaling pathways (PKA and ERK).
Conclusions:
- KINACT provides a powerful method for recording and analyzing transient kinase signaling.
- This tool enhances understanding of kinase roles in biological and disease processes.
- KINACT facilitates the discovery of novel kinase-targeted therapeutics.
- The technology aids in dissecting complex signaling networks, including oncogenic pathways.
