title: Biochemical and physiological characterization of fut4 and fut6 mutants defective in arabinogalactan-protein fucosylation in Arabidopsis.
abstract: Arabinogalactan-proteins (AGPs) are highly glycosylated hydroxyproline-rich glycoproteins present in plant cell walls. AGPs are characterized by arabinose-/galactose-rich side chains, which define their interactive molecular surface. Fucose residues are found in some dicotyledon AGPs, and AGP fucosylation is developmentally regulated. We previously identified Arabidopsis thaliana FUT4 and FUT6 genes as AGP-specific fucosyltransferases (FUTs) based on their enzymatic activities when heterologously expressed in tobacco (Nicotiana tabacum) BY2 suspension-cultured cells. Here, the functions of FUT4 and FUT6 and the physiological roles of fucosylated AGPs were further investigated using Arabidopsis fut4, fut6, and fut4/fut6 mutant plants. All mutant plants showed no phenotypic differences compared to wild-type plants under physiological conditions, but showed reduced root growth in the presence of elevated NaCl. However, roots of wild-type and fut4 mutant plants contained terminal fucose epitopes, which were absent in fut6 and fut4/fut6 mutant plants as indicated by eel lectin staining. Monosaccharide analysis showed fucose was present in wild-type leaf and root AGPs, but absent in fut4 leaf AGPs and in fut4/fut6 double mutant leaf and root AGPs, indicating that FUT4 was required for fucosylation of leaf AGPs while both FUT4 and FUT6 contributed to fucosylation of root AGPs. Glycome profiling of cell wall fractions from mutant roots and leaves showed distinct glycome profiles compared to wild-type plants, indicating that fucosyl residues on AGPs may regulate intermolecular interactions between AGPs and other wall components. The current work exemplifies the possibilities of refinement of cell wall structures by manipulation of a single or a few cell wall biosynthetic genes.
authors.affiliation: Department of Environmental and Plant Biology, Ohio University, Athens, OH 45701, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liang
authors.forename: Debarati
authors.initials: D
authors.lastname: Basu
authors.forename: Sivakumar
authors.initials: S
authors.lastname: Pattathil
authors.forename: Wen-Liang
authors.initials: WL
authors.lastname: Xu
authors.forename: Alexandra
authors.initials: A
authors.lastname: Venetos
authors.forename: Stanton L
authors.initials: SL
authors.lastname: Martin
authors.forename: Ahmed
authors.initials: A
authors.lastname: Faik
authors.forename: Michael G
authors.initials: MG
authors.lastname: Hahn
authors.forename: Allan M
authors.initials: AM
authors.lastname: Showalter
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism; D005647:Fucosyltransferases / Q000235:genetics* / Q000378:metabolism; D006031:Glycosylation; D009088:Mucoproteins / Q000378:metabolism; D009154:Mutation; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000254:growth & development / Q000276:immunology / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D009088:Mucoproteins; D010940:Plant Proteins; C006619:arabinogalactan proteins; C000590918:FUT4 protein, Arabidopsis; C000590917:FUT6 protein, Arabidopsis; D005647:Fucosyltransferases
keywords: Arabidopsis; arabinogalactan-proteins; fucosyltransferase; glycosylation; hydroxyproline-rich proteins; plant cell wall.