title: Transmembrane myosin chitin synthase involved in mollusc shell formation produced in Dictyostelium is active.
abstract: Several mollusc shells contain chitin, which is formed by a transmembrane myosin motor enzyme. This protein could be involved in sensing mechanical and structural changes of the forming, mineralizing extracellular matrix. Here we report the heterologous expression of the transmembrane myosin chitin synthase Ar-CS1 of the bivalve mollusc Atrina rigida (2286 amino acid residues, M.W. 264 kDa/monomer) in Dictyostelium discoideum, a model organism for myosin motor proteins. Confocal laser scanning immunofluorescence microscopy (CLSM), chitin binding GFP detection of chitin on cells and released to the cell culture medium, and a radiochemical activity assay of membrane extracts revealed expression and enzymatic activity of the mollusc chitin synthase in transgenic slime mold cells. First high-resolution atomic force microscopy (AFM) images of Ar-CS1 transformed cellulose synthase deficient D. discoideumdcsA(-) cell lines are shown.
authors.affiliation: INM - Leibniz Institute for New Materials, Biomineralisation Group, Campus D2.2, D-66123 Saarbrücken, Germany.
authors.forename: Veronika
authors.initials: V
authors.lastname: Schönitzer
authors.forename: Norbert
authors.initials: N
authors.lastname: Eichner
authors.forename: Hauke
authors.initials: H
authors.lastname: Clausen-Schaumann
authors.forename: Ingrid M
authors.initials: IM
authors.lastname: Weiss
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002686:Chitin / Q000096:biosynthesis; D002687:Chitin Synthase / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004023:Dictyostelium / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D049851:Gastropoda / Q000201:enzymology*; D018625:Microscopy, Atomic Force; D009218:Myosins / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000199:Actins; D011993:Recombinant Fusion Proteins; D002686:Chitin; D002687:Chitin Synthase; D009218:Myosins