title: Activation of beta-glucan synthases by wall-bound purple acid phosphatase in tobacco cells.
abstract: Wall-bound purple acid phosphatases have been shown to be potentially involved in the regulation of plant cell growth. The aim of this work was to further investigate the function of one of these phosphatases in tobacco (Nicotiana tabacum), NtPAP12, using transgenic cells overexpressing the enzyme. The transgenic cells exhibited a higher level of phosphatase activity in their walls. The corresponding protoplasts regenerating a cell wall exhibited a higher rate of beta-glucan synthesis and cellulose deposition was increased in the walls of the transgenic cells. A higher level of plasma membrane glucan synthase activities was also measured in detergent extracts of membrane fractions from the transgenic line, while no activation of Golgi-bound glycan synthases was detected. Enzymatic hydrolysis and methylation analysis were performed on the products synthesized in vitro by the plasma membrane enzymes from the wild-type and transgenic lines extracted with digitonin and incubated with radioactive UDP-glucose. The data showed that the glucans consisted of callose and cellulose and that the amount of each glucan synthesized by the enzyme preparation from the transgenic cells was significantly higher than in the case of the wild-type cells. The demonstration that callose and cellulose synthases are activated in cells overexpressing the wall-bound phosphatase NtPAP12 suggests a regulation of these carbohydrate synthases by a phosphorylation/dephosphorylation process, as well as a role of wall-bound phosphatases in the regulation of cell wall biosynthesis.
authors.affiliation: Department of Chemical and Biological Sciences, Japan Women's University, Tokyo 112-8681, Japan.
authors.forename: Rumi
authors.initials: R
authors.lastname: Kaida
authors.forename: Yumi
authors.initials: Y
authors.lastname: Satoh
authors.forename: Vincent
authors.initials: V
authors.lastname: Bulone
authors.forename: Yohko
authors.initials: Y
authors.lastname: Yamada
authors.forename: Tomomi
authors.initials: T
authors.lastname: Kaku
authors.forename: Takahisa
authors.initials: T
authors.lastname: Hayashi
authors.forename: Takako S
authors.initials: TS
authors.lastname: Kaneko
mesh_terms: D000135:Acid Phosphatase / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D002473:Cell Wall / Q000201:enzymology*; D004789:Enzyme Activation; D005110:Extracellular Space / Q000378:metabolism; D005936:Glucans / Q000378:metabolism; D005964:Glucosyltransferases / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000201:enzymology; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D011134:Polysaccharides / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D011523:Protoplasts / Q000201:enzymology; D012038:Regeneration; D013997:Time Factors; D014026:Nicotiana / Q000166:cytology* / Q000201:enzymology* / Q000235:genetics
chemical_list: D005936:Glucans; D006023:Glycoproteins; D011134:Polysaccharides; D005964:Glucosyltransferases; C478648:cellulose synthase; C091137:purple acid phosphatase; D000135:Acid Phosphatase