Role of RhoGAP Rgd1 in Pkc1 signaling-related actin repolarization under heat shock stress in Saccharomyces cerevisiae

Abstract

BackgroundA serine/threonine kinase Pkc1 is the sole protein kinase C in the budding yeast Saccharomyces cerevisiae, and plays an important role in the regulation of polarized growth and stress responses such as those due to heat shock. Exposure of cells to high temperature transiently arrests polarized growth and leads to depolarization of the actin cytoskeleton, followed by actin repolarization during adaptation to heat shock stress. Actin repolarization is ensured by the activation of Pkc1 signaling; however, the molecular mechanisms underlying this phenomenon remain poorly understood. MethodsUsing an overexpression construct of a constitutively active mutant of Pkc1 (Pkc1R398P), we explored the Pkc1 target molecules involved in actin repolarization. ResultsPKC1R398P overexpression as well as heat shock stress increased the phosphorylation levels of Rho GTPase-activating protein (RhoGAP) Rgd1. Rgd1 was found to contribute to Pkc1-signaling-related actin repolarization during adaptation to heat shock stress in a GAP activity-independent manner, with Ser148 in Rgd1 playing a crucial role. Furthermore, Rgd1 was involved in the maintenance of phosphorylation status of the mitogen-activated protein (MAP) kinase Mpk1, a downstream effector of Pkc1, under heat shock stress. ConclusionsRgd1 is a target of Pkc1 signaling under conditions of heat shock stress, and required for the normal process of actin repolarization during adaptation to heat shock stress. General significanceOur results provide insights into the molecular mechanism underlying Pkc1-mediated modulation of actin repolarization under heat shock stress.