title: Visual mapping of cell wall biosynthesis.
abstract: Biosynthesis of pectin and hemicelluloses occurs in the Golgi apparatus and is thought to involve spatial regulations and complex formation of biosynthetic enzymes and proteins. We have demonstrated that a combination of heterologous expression of recombinant proteins tagged with fluorescent proteins and live cell imaging with confocal laser scanning microscopy (CLSM) allows efficient visualization of biosynthetic enzymes and proteins in subcellular compartments. We have also successfully utilized bimolecular fluorescence complementation (BiFC) for in situ visualization of protein-protein interactions of pectin biosynthetic enzymes and for the determination of their membrane topology in the Golgi apparatus.
authors.affiliation: The Department of Plant Biology and Biotechnology, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Sakuragi
authors.forename: Morten H H
authors.initials: MH
authors.lastname: Nørholm
authors.forename: Henrik V
authors.initials: HV
authors.lastname: Scheller
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D002262:Carboxy-Lyases / Q000096:biosynthesis / Q000235:genetics; D021961:Cell Membrane Structures / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism*; D005822:Genetic Vectors; D006056:Golgi Apparatus / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D018613:Microscopy, Confocal; D010368:Pectins / Q000096:biosynthesis*; D010430:Pentosyltransferases / Q000096:biosynthesis; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014026:Nicotiana / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D014162:Transfection
chemical_list: D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010368:Pectins; D010430:Pentosyltransferases; D002262:Carboxy-Lyases; C023257:UDPglucuronate decarboxylase