Light-Mediated Control over TRPC3-Mediated NFAT Signaling
Annarita Graziani1, Bernadett Bacsa1, Denis Krivic1
1Gottfried-Schatz-Research-Center - Biophysics, Medical University of Graz, Neue Stiftingtalstrasse 6/D04, 8010 Graz, Austria.
Cells
|March 4, 2020
Summary
Researchers developed a light-controlled system to precisely regulate nuclear factor of activated T-cells (NFAT) activity using TRPC3 channels. This photopharmacological approach offers new insights into TRPC-NFAT signaling pathways.
Area of Science:
- Ion Channel Physiology
- Molecular Cell Biology
- Calcium Signaling
Background:
- Canonical transient receptor potential (TRPC) channels are implicated in pathological remodeling.
- Nuclear factor of activated T-cells (NFAT) transcription factors are downstream targets of TRPC-mediated calcium (Ca2+) signaling.
- Previous attempts to reconstitute TRPC-NFAT signaling showed limited responsiveness, suggesting a need for precise spatiotemporal control.
Purpose of the Study:
- To develop a light-inducible system for precise control of NFAT1 nuclear translocation.
- To investigate the role of TRPC3 channel activity in NFAT1 activation.
- To explore the spatiotemporal dynamics of Ca2+ signals regulating NFAT1.
Main Methods:
- Utilized a novel photochromic TRPC benzimidazole activator (OptoBI-1) and a modified TRPC3 mutant.
- Employed human embryonic kidney 293 (HEK293) cells for heterologous expression.
- Developed an all-optical protocol to monitor Ca2+ patterns and NFAT1 nuclear translocation upon light stimulation.
Main Results:
- Demonstrated light-mediated control of NFAT1 nuclear translocation using the OptoBI-1 activator and TRPC3 mutant.
- Revealed that wild-type TRPC3 exhibits constitutive NFAT1 nuclear translocation.
- Showed that a TRPC3 mutant lacking basal activity allows for spatiotemporally precise photopharmacological control of NFAT1 activity.
- Established a link between global cellular Ca2+ signals and TRPC3-mediated NFAT1 nuclear translocation.
Conclusions:
- Photopharmacology provides precise spatiotemporal control over TRPC3-NFAT signaling.
- TRPC3 activity is tightly linked to NFAT1 nuclear translocation, modulated by global Ca2+ signals.
- This system offers a novel tool to study TRPC channel function and calcium signaling in cellular processes.
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