Research Article: ROP GTPase-Dependent Actin Microfilaments Promote PIN1 Polarization by Localized Inhibition of Clathrin-Dependent Endocytosis

Date Published: April 3, 2012

Publisher: Public Library of Science

Author(s): Shingo Nagawa, Tongda Xu, Deshu Lin, Pankaj Dhonukshe, Xingxing Zhang, Jiri Friml, Ben Scheres, Ying Fu, Zhenbiao Yang, Mark Estelle

Abstract: A study in leaf epidermal pavement cells reveals that auxin activation of a Rho-like GTPase from plants induces inhibition of endocytosis through the clathrin-mediated pathway by regulating the accumulation of cortical F-actin.

Partial Text: Cell polarity is a conserved cellular property that is necessary for the generation of diverse forms and types of cells in both uni- and multicellular organisms [1],[2]. The general design principles that govern the formation of polarity and how they are used to generate diverse forms of polarity is a fundamental issue of developmental mechanisms. In the unicellular yeast, Rho family GTPase-mediated activation of endocytosis is required for cell polarization [3]–[5]. In contrast, emerging evidence suggests that Rho family GTPase-mediated inhibition of endocytosis is essential for the polarization of cells in some multicellular tissues as shown in cultured epithelial cells from rat [6] and neuroectodermal epithelial cells from Drosophila[7]. It is unclear whether Rho-mediated inhibition of endocytosis is a common design principle for polarity establishment in multicellular systems and how the inhibition of endocytosis is regulated.

Our findings here have established an auxin-activated ROP2-signaling pathway that regulates PIN1 protein polarization to the PC lobe through the localized inhibition of PIN protein endocytosis. Given the requirement of PIN1 for the ROP2 activation at the lobe region of the PM [19], this signaling pathway underscores a positive feedback loop leading to PIN1 polarization, which provides strong support for the hypothesis that auxin acts as a self-organizing signal in the control of PIN-dependent auxin efflux [14],[15]. Furthermore, we have demonstrated that auxin signaling links the Rho GTPase-dependent accumulation of the cortical fine F-actin to PIN polarization. This finding provides an important insight into the mechanism for the modulation of F-actin reorganization in its regulation of PIN endocytosis and polarization [32],[47],[49]. Several recent studies implicate actin dynamics in the regulation of PIN endocytic recycling. By using transgenic rice plants that express different levels of mouse talin protein, Nick et al. recently showed that dynamics of actin organization and auxin transport efficiency are coupled [34]. Auxin transport inhibitors such as TIBA were shown to induce bundling of actin filaments and inhibit endocytosis, and thus were suggested to affect auxin transport through actin-mediated vesicle trafficking of auxin transport-related proteins [13]. Our data show that the ROP2/RIC4-dependent auxin signaling pathway induces the accumulation of the cortical fine F-actin, which inhibits clathrin-dependent PIN1 endocytosis that leads to PIN1 polarization. The mechanism by which the ROP2-dependent F-actin accumulation inhibits endocytosis needs to be investigated in the future. In yeast, clathrin-dependent endocytosis requires not only Cdc42 GTPase-dependent polymerization of cortical actin patches but also their dynamics. Similarly in pollen tubes both ROP1 GTPase-dependent polymerization and dynamics of tip F-actin are critical for polarized pollen tube growth [50],[51]. Thus it will be interesting to know whether the polymerization of ROP2-mediated F-actin is also important for clathrin-dependent PIN1 endocytosis.