A CD317/tetherin-RICH2 complex plays a critical role in the organization of the subapical actin cytoskeleton in polarized epithelial cells.
CD317/tetherin is a lipid raft-associated integral membrane protein with a novel topology. It has a short N-terminal cytosolic domain, a conventional transmembrane domain, and a C-terminal glycosyl-phosphatidylinositol anchor. We now show that CD317 is expressed at the apical surface of polarized epithelial cells, where it interacts indirectly with the underlying actin cytoskeleton. CD317 is linked to the apical actin network via the proteins RICH2, EBP50, and ezrin. Knocking down expression of either CD317 or RICH2 gives rise to the same phenotype: a loss of the apical actin network with concomitant loss of apical microvilli, an increase in actin bundles at the basal surface, and a reduction in cell height without any loss of tight junctions, transepithelial resistance, or the polarized targeting of apical and basolateral membrane proteins. Thus, CD317 provides a physical link between lipid rafts and the apical actin network in polarized epithelial cells and is crucial for the maintenance of microvilli in such cells.
Rollason
Ruth
R
Department of Biochemistry, University of Bristol, Bristol BS8 1TD, England, UK.
Korolchuk
Viktor
V
Hamilton
Clare
C
Jepson
Mark
M
Banting
George
G
D008841:Actin Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D019556:COS Cells; D018938:Caco-2 Cells; D016764:Cell Polarity / Q000502:physiology*; D002522:Chlorocebus aethiops; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D015536:Down-Regulation / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D058851:GPI-Linked Proteins; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D020744:rac GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000099149:Ezrin; D000074010:Bone Marrow Stromal Antigen 2
D000199:Actins; D015703:Antigens, CD; D003598:Cytoskeletal Proteins; D058851:GPI-Linked Proteins; D020690:GTPase-Activating Proteins; D008562:Membrane Glycoproteins; D034741:RNA, Small Interfering; D020744:rac GTP-Binding Proteins; D020741:rho GTP-Binding Proteins; D000099149:Ezrin; C539906:ARHGAP44 protein, human; C094572:BST2 protein, human; D000074010:Bone Marrow Stromal Antigen 2