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bioRxiv · 10.1101/2024.03.06.583600

Cooperative involvement of zipper-interacting protein kinase (ZIPK) and the dual-specificity cell division cycle 14A phosphatase (CDC14A) in vascular smooth muscle cell migration

Al-Ghabkari, A.·Carlson, D. A.·Haystead, T. A.·MacDonald, J. A.
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Abstract

Zipper-interacting protein kinase (ZIPK) is a Ser/Thr protein kinase with regulatory involvement in vascular smooth muscle cell (VSMC) actin polymerization and focal adhesion assembly dynamics. ZIPK silencing can induce cytoskeletal remodeling with disassembly of actin stress fiber networks and coincident loss of focal adhesion kinase (FAK)-pY397 phosphorylation. The link between ZIPK inhibition and FAK phosphorylation is unknown, and critical interactor(s) and regulator(s) are not yet defined. In this study, we further analyzed the ZIPK-FAK relationship in VSMCs. The application of HS38, a selective ZIPK inhibitor, to coronary artery vascular smooth muscle cells (CASMCs) suppressed cell migration, myosin light chain phosphorylation (pT18&pS19) and FAK-pY397 phosphorylation as well. This was associated with the translocation of cytoplasmic FAK to the nucleus. ZIPK inhibition with HS38 was consistently found to suppress the activation of FAK and attenuate the phosphorylation of other focal adhesion protein components (i.e., pCas130, paxillin, ERK). In addition, our study showed a decrease in human cell-division cycle 14A phosphatase (CDC14A) levels with ZIPK-siRNA treatment and increased CDC14A with transient transfection of ZIPK. Proximity ligation assays (PLA) revealed CDC14A localized with ZIPK and FAK. Silencing CDC14A showed an increase of FAK-pY397 phosphorylation. Ultimately, the data presented herein strongly support a regulatory mechanism of FAK in CASMCs by a ZIPK-CDC14A partnership; ZIPK may act as a key signal integrator to control CDC14A and FAK during VSMC migration.

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Al-Ghabkari, A., Carlson, D. A., Haystead, T. A., MacDonald, J. A.. 2024-03-09. Cooperative involvement of zipper-interacting protein kinase (ZIPK) and the dual-specificity cell division cycle 14A phosphatase (CDC14A) in vascular smooth muscle cell migration. https://doi.org/10.1101/2024.03.06.583600

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