title: Concentration of an integral membrane protein, CD43 (leukosialin, sialophorin), in the cleavage furrow through the interaction of its cytoplasmic domain with actin-based cytoskeletons.
abstract: In leukocytes such as thymocytes and basophilic leukemia cells, a glycosilated integral membrane protein called CD43 (leukosialin or sialophorin), which is defective in patients with Wiskott-Aldrich syndrome, was highly concentrated in the cleavage furrow during cytokinesis. Not only at the mitotic phase but also at interphase, CD43 was precisely colocalized with ezrin-radixin-moesin family members. (ERM), which were previously reported to play an important role in the plasma membrane-actin filament association in general. At the electron microscopic level, throughout the cell cycle, both CD43 and ERM were tightly associated with microvilli, providing membrane attachment sites for actin filaments. We constructed a cDNA encoding a chimeric molecule consisting of the extracellular domain of mouse E-cadherin and the transmembrane/cytoplasmic domain of rat CD43, and introduced it into mouse L fibroblasts lacking both endogenous CD43 and E-cadherin. In dividing transfectants, the chimeric molecules were concentrated in the cleavage furrow together with ERM, and both proteins were precisely colocalized throughout the cell cycle. Furthermore, using this transfection system, we narrowed down the domain responsible for the CD43-concentration in the cleavage furrow. Based on these findings, we conclude that CD43 is concentrated in the cleavage furrow through the direct or indirect interaction of its cytoplasmic domain with ERM and actin filaments.
authors.affiliation: Department of Information Physiology, National Institute for Physiological Sciences, Aichi, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Yonemura
authors.forename: A
authors.initials: A
authors.lastname: Nagafuchi
authors.forename: N
authors.initials: N
authors.lastname: Sato
authors.forename: S
authors.initials: S
authors.lastname: Tsukita
mesh_terms: D000199:Actins / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD; D001483:Base Sequence; D015820:Cadherins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D007739:L Cells; D051917:Leukosialin; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology / Q000648:ultrastructure; D051379:Mice; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism; D012795:Sialoglycoproteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D000199:Actins; D015703:Antigens, CD; D015820:Cadherins; D051917:Leukosialin; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; C497863:Spn protein, mouse; C497864:Spn protein, rat