Auxin-induced nanoclustering of membrane signaling complexes underlies cell polarity establishment in Arabidopsis

Cell polarity is fundamental to the development of both eukaryotic and prokaryotic organisms, yet the mechanism of its establishment remains poorly understood. Here we show that signal-activated nanoclustering of membrane proteins and a cytoskeleton-based feedback loop provide an important mechanism for the establishment of cell polarity. The phytohormone auxin promoted sterol-dependent nanoclustering of cell surface transmembrane receptor-like kinase 1 (TMK1) to initiate cell polarity during the morphogenesis of Arabidopsis puzzle piece-shaped leaf pavement cells (PC). Auxin-triggered nanoclustering of TMK1 stabilized flotillin-associated ordered nanodomains, which were essential for auxin-mediated formation of ROP6 GTPase nanoclusters that act downstream TMK1 to promote cortical microtubule ordering. Mathematical modeling further demonstrated the essential role of this auxin-mediated stabilization of TMK1 and ROP6 nanoclusters, and predicted the additional requirement of ROP6-dependent cortical microtubules for further stabilization of TMK1-sterol nanodomains and the polarization of PC. This prediction was experimentally validated by genetic and biochemical data. Our studies reveal a new paradigm for polarity establishment: A diffusive signal triggers cell polarization by activating cell surface receptor-mediated lateral segregation of signaling components and a cytoskeleton-mediated positive feedback loop of nanodomain stabilization.\n\nHighlightsO_LISterols are required for cell polarity in Arabidopsis leaf epidermal cells\nC_LIO_LIAuxin promotes lipid ordering and polar distribution of ordered lipid nanodomains at the plasma membrane (PM)\nC_LIO_LIAuxin stabilizes sterol-dependent nanoclustering of transmembrane kinase (TMK1), a PM auxin signal transducer\nC_LIO_LIAuxin-induced TMK1 nanoclustering is required but insufficient for cell polarization\nC_LIO_LIMicrotubule-based feedback stabilization of the auxin-induced TMK1 nanodomains can generate cell polarity\nC_LI