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Quantitative analysis reveals that actin and Src-family kinases regulate nuclear YAP1 and its export
journal contributionposted on 2020-07-15, 11:09 authored by Nil Ege, Anna M Dowbaj, Ming Jiang, Michael Howell, Steven Hooper, Charles Foster, Robert P Jenkins, Erik Sahai
The transcriptional regulator YAP1 is critical for the pathological activation of fibroblasts. In normal fibroblasts, YAP1 is located in the cytoplasm, while in activated cancer-associated fibroblasts, it is nuclear and promotes the expression of genes required for pro-tumorigenic functions. Here, we investigate the dynamics of YAP1 shuttling in normal and activated fibroblasts, using EYFP-YAP1, quantitative photobleaching methods, and mathematical modeling. Imaging of migrating fibroblasts reveals the tight temporal coupling of cell shape change and altered YAP1 localization. Both 14-3-3 and TEAD binding modulate YAP1 shuttling, but neither affects nuclear import. Instead, we find that YAP1 nuclear accumulation in activated fibroblasts results from Src and actomyosin-dependent suppression of phosphorylated YAP1 export. Finally, we show that nuclear-constrained YAP1, upon XPO1 depletion, remains sensitive to blockade of actomyosin function. Together, these data place nuclear export at the center of YAP1 regulation and indicate that the cytoskeleton can regulate YAP1 within the nucleus.
Src-family kinasesYAP1actomyosin cytoskeletonfibroblastmathematical modelingnuclear exportphotobleachingActinsActive Transport, Cell NucleusAdaptor Proteins, Signal TransducingAnimalsCell Cycle ProteinsCell Line, TumorCell MovementCell NucleusCell ProliferationCytoplasmCytoskeletonDNA-Binding ProteinsFibroblastsGene Expression RegulationHumansMiceModels, TheoreticalPhosphoproteinsPhosphorylationPhotobleachingSignal Transductionsrc-Family KinasesSahai FC001144Treisman FC001190HTS