title: Characterization of moesin in the sea urchin Lytechinus variegatus: redistribution to the plasma membrane following fertilization is inhibited by cytochalasin B.
abstract: We have investigated the distribution and function of an ezrin-radixin-moesin-like (ERM) molecule in the sea urchin. A sea urchin homologue of moesin was cloned that shares 75% amino acid similarity in the conserved N-terminal region to other moesin molecules. A 6.3 kb message is transcribed late in embryogenesis and is present in adult tissues. Polyclonal antibodies were generated to proteins expressed by a bacterial expression vector, and affinity purified. These antibodies recognize a single 75 kDa protein that is present throughout development in approximately equal abundance, and specifically they immuno-precipitate a single protein. We show by immunolocalization that SUmoesin has two predominant patterns during development. First, SUmoesin is rapidly redistributed after fertilization from a location throughout the egg cytoplasm to a location in the egg cortex. Later in embryogenesis, SUmoesin is localized to the apical ends of cells in the regions of cell-cell junctions. We show that SUmoesin is present in actin-rich regions of the embryo. Finally, we show that the location of SUmoesin requires an intact actin-based cytoskeleton. SUmoesin fails to localize to the plasma membrane after fertilization in the presence of cytochalasin B. Furthermore, SUmoesin loses its apical position in the region of cell-cell junctions in the presence of cytochalasin B in later stages of embryogenesis. This effect is reversible, and the microtubule inhibitor colchicine has no effect. These results show that SUmoesin becomes associated with apical plasma membrane structures early in development, and that SUmoesin is both coincident with actin and requires the assembly of actin filaments to maintain its localization.
authors.affiliation: Duke University, Durham, NC 27708, USA.
authors.forename: E S
authors.initials: ES
authors.lastname: Bachman
authors.forename: D R
authors.initials: DR
authors.lastname: McClay
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism / Q000502:physiology*; D003001:Cloning, Molecular; D003078:Colchicine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D004625:Embryo, Nonmammalian / Q000502:physiology*; D005260:Female; D005306:Fertilization; D005775:Gastrula / Q000502:physiology*; D006801:Humans; D008297:Male; D051379:Mice; D008840:Microfilament Proteins; D008969:Molecular Sequence Data; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification; D012617:Sea Urchins; D017386:Sequence Homology, Amino Acid; D013094:Spermatozoa / Q000502:physiology
chemical_list: D008840:Microfilament Proteins; D011506:Proteins; D011994:Recombinant Proteins; C071058:moesin; D003571:Cytochalasin B; D003078:Colchicine