Article
Version 1
Preserved in Portico This version is not peer-reviewed
Crosstalk among WEE1 Kinase, AKT, and GSK3 in Nav1.2 Channelosome Regulation
Version 1
: Received: 2 June 2024 / Approved: 3 June 2024 / Online: 3 June 2024 (08:07:35 CEST)
How to cite: Singh, A. K.; Singh, J.; Laezza, F. Crosstalk among WEE1 Kinase, AKT, and GSK3 in Nav1.2 Channelosome Regulation. Preprints 2024, 2024060034. https://doi.org/10.20944/preprints202406.0034.v1 Singh, A. K.; Singh, J.; Laezza, F. Crosstalk among WEE1 Kinase, AKT, and GSK3 in Nav1.2 Channelosome Regulation. Preprints 2024, 2024060034. https://doi.org/10.20944/preprints202406.0034.v1
Abstract
The signaling complex around voltage-gated sodium (Nav) channels includes accessory proteins and kinases crucial for regulating neuronal firing. Previous studies showed that one such kinase, WEE1—critical to the cell cycle—selectively modulates Nav1.2 channel activity through the accessory protein fibroblast growth factor 14 (FGF14). Here, we tested whether WEE1 exhibits crosstalk with the AKT/GSK3 pathway for coordinated regulation of FGF14/Nav1.2 channel complex assembly and function. Using the in-cell split luciferase complementation assay (LCA), we found that the WEE1 inhibitor II and GSK3 inhibitor XIII reduce FGF14/Nav1.2 complex formation, while the AKT inhibitor triciribine increases it. However, combining WEE1 inhibitor II with either one of the other two inhibitors abolished its effect on FGF14/Nav1.2 complex formation. Whole-cell voltage-clamp recordings of sodium currents (INa) in HEK293 cells co-expressing Nav1.2 channels and FGF14-GFP showed that WEE1 inhibitor II significantly suppresses peak INa density, both alone and in the presence of triciribine or GSK3 inhibitor XIII, despite the latter inhibitors’ opposite effects on INa. Additionally, WEE1 inhibitor II slowed the tau of fast inactivation, and caused depolarizing shifts in the voltage dependence of activation and inactivation. These phenotypes either prevailed or were additive when combined with triciribine but were outcompeted when both WEE1 inhibitor II and GSK3 inhibitor XIII were present. Concerted regulation by WEE1 inhibitor II, triciribine and GSK3 inhibitor XIII were also observed on long-term inactivation and use-dependence of Nav1.2 currents. Overall, these findings suggest a complex role for WEE1 kinase—in concert with the AKT/GSK3 pathway—in regulating the Nav1.2 channelosome.
Keywords
voltage-gated sodium channels; fibroblast growth factors; split luciferase assay; kinase inhibitors
Subject
Biology and Life Sciences, Neuroscience and Neurology
Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Comments (0)
We encourage comments and feedback from a broad range of readers. See criteria for comments and our Diversity statement.
Leave a public commentSend a private comment to the author(s)
* All users must log in before leaving a comment
