title: Anthrax toxin receptor 1/tumor endothelium marker 8 mediates cell spreading by coupling extracellular ligands to the actin cytoskeleton.
abstract: Tumor endothelial marker 8 (TEM8) is induced in tumor-associated vasculature and acts as a receptor for Protective Antigen (PA), the cell-binding component of the anthrax toxin determinant for toxin entrance into cells. However, the normal function for TEM8 remains unknown. We show that TEM8 functions as an adhesion molecule mediating cell spreading on immobilized PA and collagen I. The mechanism for TEM8 interaction with collagen I was cell type-specific, because binding to collagen I was abrogated by beta1 integrin function blocking antibody in HEK293 cells, but not in primary synovial rabbit fibroblasts. Binding to PA remained unaffected by the addition of beta1 integrin function blocking antibody. Whereas the extracellular and transmembrane domains of TEM8 were sufficient to provide cell attachment, the intracellular domain was critical for spreading. Fusion of the cytosolic domain of TEM8 to the IL-2 receptor, conferred cell-spreading capability on IL-2 receptor antibody substrates. The cytoplasmic domain mediated linkage with the actin cytoskeleton as it co-precipitated actin and determined partitioning of TEM8 to the actin-containing detergent insoluble cellular fraction. TEM8 anchorage to actin was relevant as spreading was inhibited by the cytoskeleton-disrupting drug cytochalasin D, but persisted in the presence of the microtubule-depolymerizing drug nocodazole, and in cells lacking intermediate filaments. Thus, our results indicate that TEM8 is a new adhesion molecule linking collagen I or PA to the actin cytoskeleton.
authors.affiliation: Department of Cell Biology , Emory University, Atlanta, Georgia 30322-3030, USA. ericaw@cellbio.emory.edu
authors.forename: Erica
authors.initials: E
authors.lastname: Werner
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Kowalczyk
authors.forename: Victor
authors.initials: V
authors.lastname: Faundez
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry; D001427:Bacterial Toxins / Q000737:chemistry; D002448:Cell Adhesion; D024042:Collagen Type I / Q000737:chemistry; D003599:Cytoskeleton / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology*; D008840:Microfilament Proteins; D009363:Neoplasm Proteins / Q000378:metabolism / Q000502:physiology*; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000378:metabolism / Q000502:physiology*
chemical_list: C473287:ANTXR1 protein, human; D000199:Actins; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D024042:Collagen Type I; D003902:Detergents; D008024:Ligands; D008565:Membrane Proteins; D008840:Microfilament Proteins; D009363:Neoplasm Proteins; D011956:Receptors, Cell Surface; C030325:anthrax toxin