Protein Kinase A activity is regulated by actomyosin contractility during cell migration and is required for durotaxis

Abstract

Dynamic subcellular regulation of Protein kinase A (PKA) activity is important for the motile behavior of many cell types, yet the mechanisms governing PKA activity during cell migration remain largely unknown. The motility of SKOV-3 epithelial ovarian cancer (EOC) cells has been shown to be dependent on both localized PKA activity and, more recently, on mechanical reciprocity between cellular tension and extracellular matrix (ECM) rigidity. Here, we investigated the possibility that PKA is regulated by mechanical signaling during migration. We find that localized PKA activity in migrating cells rapidly decreases upon inhibition of actomyosin contractility (specifically, of myosin ATPase, ROCK (Rho kinase), or MLCK (myosin light chain kinase) activity). Moreover, PKA activity is spatially and temporally correlated with cellular traction forces in migrating cells. Additionally, PKA is rapidly and locally activated by mechanical stretch in an actomyosin contractility-dependent manner. Finally, inhibition of PKA activity inhibits mechanicallyguided migration, also known as durotaxis. These observations establish PKA as a locallyregulated effector of cellular mechanotransduction and as a regulator of mechanicallyguided cell migration.All rights reserved. No reuse allowed without permission.

All Roads Lead to Directional Cell Migration

“…Through these signalling networks, effector proteins like PKA and YAP become activated, leading to changes in protein activation and dynamics, as well as gene expression changes, which are crucial for normal durotaxis [79,80].…”

“…In durotaxis, stress fibre contractility, which is normally mediated by RhoA, is the means by which forces are applied on the substrate [28]. Active RhoA also controls fast cellular retraction during durotaxis [79]. These activities of RhoA are also evident and required for haptotaxis and chemotaxis [109,110].…”

Multiphysics simulation of a compression–perfusion combined bioreactor to predict the mechanical microenvironment during bone metastatic breast cancer loading experiments

“…In a novel microfluidic device, when mechanically flow osteocytes were adjacent to breast cancer cells, breast cancer extravasation was significantly reduced with mechanically stimulated osteocytes compared to static osteocytes, though the breast cancer cells remained under static conditions (Mei et al, 2019). When considering the effects of strain, 2D stretching is typically applied acutely and held steady for a period of time with a variety of results across multiple cell types (Gao & Carson, 2016; Manome et al, 2003; McKenzie et al, 2020; Panzetta et al, 2019; Riching et al, 2014). One study reported that 2D cyclic compression of breast cancer cells plated underneath an agarose gel by a platen regulated necrosis vs apoptosis, and the mode of death was sensitive to loading frequency, peak applied compressive displacement, and duration of loading bout (Takao et al, 2019).…”

DrFLINC Contextualizes Super-resolution Activity Imaging

“…We performed two-color pcSOFI with DrFLINC-AKAR and Lifeact-Dronpa in HeLa cells and found that with the exception of filopodia, the majority of PKA activity microdomains did not colocalize with actin (Figure 4B/E). These data suggest that PKA activity microdomains are not generally organized along actin networks, but instead more specifically targeted to sub-regions that are involved in migration and mechano-sensing [21][22] .…”