Abstract
PKA contributes to many physiological processes, including glucose homeostasis and cell migration. The substrate specificity of PKA is low compared with other kinases; thus, complex formation with A-kinase-anchoring proteins is important for the localization of PKA in specific subcellular regions and the phosphorylation of specific substrates. Here, we show that PKA forms a complex with WAVE2 (Wiskott-Aldrich syndrome protein family verprolin-homologous protein 2) in MDA-MB-231 breast cancer cells and mouse brain extracts. Two separate regions of WAVE2 are involved in WAVE2-PKA complex formation. This complex localizes to the leading edge of MDA-MB-231 cells. PKA activation results in enlargement of the membrane protrusion. WAVE2 depletion impairs PKA localization at membrane protrusions and the enlargement of membrane protrusion induced by PKA activation. Together, these results suggest that WAVE2 works as an A-kinase-anchoring protein that recruits PKA at membrane protrusions and plays a role in the enlargement of membrane protrusions induced by PKA activation.
Highlights
WASP family proteins link various upstream signals to the stimulation of Arp2/3-mediated actin polymerization
These results suggest that WAVE2 works as an A-kinase-anchoring proteins (AKAPs) that recruits PKA at membrane protrusions and plays a role in the enlargement of membrane protrusions induced by PKA activation
AKAPs, as well as other proteins associated with PKA that contribute to PKA-mediated cell migration, are not fully understood
Summary
WASP family proteins link various upstream signals to the stimulation of Arp2/3-mediated actin polymerization. To further confirm the association, endogenously expressed WAVE2 was immunoprecipitated from MDA-MB-231 breast cancer cells with anti-WAVE2 antibody, and coprecipitated PKA was examined by immunoblotting with anti-PKA C subunit antibody. The observation that holoenzymes, as well as both C and R subunits, can associate with WAVE2 directly is consistent with the results of Fig. 1 (E and F), in which WAVE2 partially dissociated from the PKA C subunit after treatment with 8-bromo-cAMP. PKA Colocalizes with WAVE2 at the Leading Edge of Membrane Protrusions—WAVE2 regulates membrane protrusions and cell migration by promoting Arp2/3 complex-dependent actin reorganization.
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