Light-gated integrator for highlighting kinase activity in living cells

Wei Lin; Abhishek Phatarphekar; Yanghao Zhong; Longwei Liu; Hyung-Bae Kwon; William H Gerwick; Yingxiao Wang; Sohum Mehta; Jin Zhang

doi:10.1038/s41467-024-51270-4

Light-gated integrator for highlighting kinase activity in living cells

Nat Commun. 2024 Sep 6;15(1):7804. doi: 10.1038/s41467-024-51270-4.

Authors

Wei Lin¹, Abhishek Phatarphekar², Yanghao Zhong^{2

3}, Longwei Liu⁴, Hyung-Bae Kwon⁵, William H Gerwick⁶, Yingxiao Wang⁴, Sohum Mehta², Jin Zhang^{7

8

9

10}

Affiliations

¹ Department of Pharmacology, University of California San Diego, La Jolla, CA, USA. wel328@health.ucsd.edu.
² Department of Pharmacology, University of California San Diego, La Jolla, CA, USA.
³ Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, USA.
⁴ Alfred E. Mann Department of Biomedical Engineering, University of Southern California, Los Angeles, CA, USA.
⁵ Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁶ Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography and Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, USA.
⁷ Department of Pharmacology, University of California San Diego, La Jolla, CA, USA. jzhang32@health.ucsd.edu.
⁸ Moores Cancer Center, University of California San Diego, La Jolla, CA, USA. jzhang32@health.ucsd.edu.
⁹ Shu Chien - Gene Lay Department of Bioengineering, University of California San Diego, La Jolla, CA, USA. jzhang32@health.ucsd.edu.
¹⁰ Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA. jzhang32@health.ucsd.edu.

Abstract

Protein kinases are key signaling nodes that regulate fundamental biological and disease processes. Illuminating kinase signaling from multiple angles can provide deeper insights into disease mechanisms and improve therapeutic targeting. While fluorescent biosensors are powerful tools for visualizing live-cell kinase activity dynamics in real time, new molecular tools are needed that enable recording of transient signaling activities for post hoc analysis and targeted manipulation. Here, we develop a light-gated kinase activity coupled transcriptional integrator (KINACT) that converts dynamic kinase signals into "permanent" fluorescent marks. KINACT enables robust monitoring of kinase activity across scales, accurately recording subcellular PKA activity, highlighting PKA activity distribution in 3D cultures, and identifying PKA activators and inhibitors in high-throughput screens. We further leverage the ability of KINACT to drive signaling effector expression to allow feedback manipulation of the balance of Gα_s^R201C-induced PKA and ERK activation and dissect the mechanisms of oncogenic G protein signaling.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Biosensing Techniques / methods
Cyclic AMP-Dependent Protein Kinases* / metabolism
HEK293 Cells
Humans
Light
Signal Transduction*

Light-gated integrator for highlighting kinase activity in living cells

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