@article{da8fe94143074f50846dad41605a0521,
title = "Integrin Mechano-chemical Signaling Generates Plasma Membrane Nanodomains that Promote Cell Spreading",
abstract = "Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are a major class of lipid-anchored plasma membrane proteins. GPI-APs form nanoclusters generated by cortical acto-myosin activity. While our understanding of the physical principles governing this process is emerging, the molecular machinery and functional relevance of GPI-AP nanoclustering are unknown. Here, we first show that a membrane receptor signaling pathway directs nanocluster formation. Arg-Gly-Asp motif-containing ligands bound to the β1-integrin receptor activate src and focal adhesion kinases, resulting in RhoA signaling. This cascade triggers actin-nucleation via specific formins, which, along with myosin activity, drive the nanoclustering of membrane proteins with actin-binding domains. Concurrently, talin-mediated activation of the mechano-transducer vinculin is required for the coupling of the acto-myosin machinery to inner-leaflet lipids, thereby generating GPI-AP nanoclusters. Second, we show that these nanoclusters are functional; disruption of their formation either in GPI-anchor remodeling mutants or in vinculin mutants impairs cell spreading and migration, hallmarks of integrin function.",
keywords = "GPI-anchored proteins, active actin-membrane composite, active rafts, cell signaling, cell spreading, integrin, mechanotransduction, membrane domains, nanoclusters, vinculin",
author = "Kalappurakkal, {Joseph Mathew} and Anilkumar, {Anupama Ambika} and Chandrima Patra and {van Zanten}, {Thomas S.} and Sheetz, {Michael P.} and Satyajit Mayor",
note = "Funding Information: We thank Cheng-han Yu for help with standardizing the SLB system; the Nano and Microfabrication Core at the Mechanobiology Institute, NUS, Singapore for providing the chromium nanopatterned surfaces; Kabir Husain and Balaji Ramalingam for MATLAB codes; Taroh Kinoshita, Yusuke Maeda, Daniel Rosel, Clare M. Waterman, and Ana Pasapera for their generous gifts of various reagents; Max Planck-NCBS Lipid center and Bini Ramachandran for mass spectrometry; the protein purification services offered at C-CAMP; and the Central Imaging and Flow Cytometry Facility (NCBS). We thank Madan Rao and Subhasri Ghosh for inspiration and Mayor laboratory members and Muriel Grammont for their valuable comments on the manuscript. J.M.K. and C.P acknowledge pre-doctoral fellowships from NCBS-TIFR. A.A.A. acknowledges a N-PDF fellowship from DST-SERB (Government of India). T.S.v.Z. acknowledges an EMBO fellowship (ALTF 1519-2013) and a NCBS Campus fellowship. S.M. acknowledges a JC Bose Fellowship from DST (Government of India), support from the NCBS-Max Planck Lipid Centre, a grant from HFSP RGP0027/2012, and support from Wellcome Trust/DBT India Alliance Margdarshi Fellowship (IA/M/15/1/502018). J.M.K. A.A.A. and S.M. designed the study. C.P performed the lipid ordering and filipin-labeling experiments. T.S.v.Z. performed and analyzed the FCS data and analyzed the anisotropy and area changes during cell spreading. The SLB experiments were standardized by J.M.K working in collaboration with M.P.S.{\textquoteright}s laboratory. J.M.K. and A.A.A. set up the EA-TIRFM microscope, performed all other experiments, and analyzed the data. J.M.K. A.A.A. and S.M. drafted the manuscript with input from all the authors. The authors declare no competing interests. Funding Information: We thank Cheng-han Yu for help with standardizing the SLB system; the Nano and Microfabrication Core at the Mechanobiology Institute, NUS, Singapore for providing the chromium nanopatterned surfaces; Kabir Husain and Balaji Ramalingam for MATLAB codes; Taroh Kinoshita, Yusuke Maeda, Daniel Rosel, Clare M. Waterman, and Ana Pasapera for their generous gifts of various reagents; Max Planck-NCBS Lipid center and Bini Ramachandran for mass spectrometry; the protein purification services offered at C-CAMP; and the Central Imaging and Flow Cytometry Facility (NCBS). We thank Madan Rao and Subhasri Ghosh for inspiration and Mayor laboratory members and Muriel Grammont for their valuable comments on the manuscript. J.M.K. and C.P acknowledge pre-doctoral fellowships from NCBS-TIFR . A.A.A. acknowledges a N-PDF fellowship from DST-SERB (Government of India). T.S.v.Z. acknowledges an EMBO fellowship ( ALTF 1519-2013 ) and a NCBS Campus fellowship. S.M. acknowledges a JC Bose Fellowship from DST (Government of India), support from the NCBS-Max Planck Lipid Centre , a grant from HFSP RGP0027/2012 , and support from Wellcome Trust/DBT India Alliance Margdarshi Fellowship ( IA/M/15/1/502018 ). Publisher Copyright: {\textcopyright} 2019 The Authors",
year = "2019",
month = jun,
day = "13",
doi = "10.1016/j.cell.2019.04.037",
language = "English (US)",
volume = "177",
pages = "1738--1756.e23",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "7",
}