Light-gated integrator for highlighting kinase activity in living cells.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 39242543.
- Also identified by DOI 10.1038/s41467-024-51270-4 and PMC identifier 11379911.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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<sub>s</sub><sup>R201C</sup>-induced PKA and ERK activation and dissect the mechanisms of oncogenic G protein signaling.
Medical subject headings
- Cyclic AMP-Dependent Protein Kinases
- Signal Transduction