Proteomic and biochemical evidence links the callose synthase in <i>Nicotiana alata</i> pollen tubes to the product of the <i>NaGSL1</i> gene

Brownfield, Lynette; Ford, Kris; Doblin, Monika S.; Newbigin, Ed; Read, Stephen; Bacic, Antony

doi:10.1111/j.1365-313x.2007.03219.x

Cited by 58 publications

(48 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The GSL genes encode (1,3)-␤-D-glucan synthases (callose synthases) (7,8), and the CESA genes encode cellulose synthases (9). The CSLs are believed to encode enzymes that synthesize the backbone of various non-cellulosic ␤-linked polysaccharides of the wall (10)(11)(12)(13).…”

mentioning

confidence: 99%

A barley cellulose synthase-like CSLH gene mediates (1,3;1,4)-β- d -glucan synthesis in transgenic Arabidopsis

Doblin

Pettolino

Wilson

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

250

288

View full text Add to dashboard Cite

The walls of grasses and related members of the Poales are characterized by the presence of the polysaccharide (1,3, 1,4)-␤-Dglucan (␤-glucan). To date, only members of the grass-specific cellulose synthase-like F (CSLF) gene family have been implicated in its synthesis. Assuming that other grass-specific CSL genes also might encode synthases for this polysaccharide, we cloned HvC-SLH1, a CSLH gene from barley (Hordeum vulgare L.), and expressed an epitope-tagged version of the cDNA in Arabidopsis, a species with no CSLH genes and no ␤-glucan in its walls. Transgenic Arabidopsis lines that had detectable amounts of the epitopetagged HvCSLH1 protein accumulated ␤-glucan in their walls. The presence of ␤-glucan was confirmed by immunoelectron microscopy (immuno-EM) of sectioned tissues and chemical analysis of wall extracts. In the chemical analysis, characteristic tri-and tetra-saccharides were identified by high-performance anion-exchange chromatography and MALDI-TOF MS following their release from transgenic Arabidopsis walls by a specific ␤-glucan hydrolase. Immuno-EM also was used to show that the epitopetagged HvCSLH1 protein was in the endoplasmic reticulum and Golgi-associated vesicles, but not in the plasma membrane. In barley, HvCSLH1 was expressed at very low levels in leaf, floral tissues, and the developing grain. In leaf, expression was highest in xylem and interfascicular fiber cells that have walls with secondary thickenings containing ␤-glucan. Thus both the CSLH and CSLF families contribute to ␤-glucan synthesis in grasses and probably do so independently of each other, because there is no significant transcriptional correlation between these genes in the barley tissues surveyed.

show abstract

mentioning

confidence: 99%

A barley cellulose synthase-like CSLH gene mediates (1,3;1,4)-β- d -glucan synthesis in transgenic Arabidopsis

Doblin

Pettolino

Wilson

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

250

288

View full text Add to dashboard Cite

show abstract

“…It was hypothesized that the same enzyme may be involved in the biosynthesis of callose and cellulose and that the biosynthesis of either polysaccharide may be controlled by phosphorylation (Delmer, 1999). However, experimental data showed that the cellulose and callose synthase catalytic subunits are encoded by different genes (Pear et al, 1996;Arioli et al, 1998;Cui et al, 2001;Doblin et al, 2001;Hong et al, 2001a;Li et al, 2003;Brownfield et al, 2007). Indeed, these proteins show no significant sequence similarity and are grouped in different glycosyltransferase families in the CAZY database (http:// www.cazy.org), family 2 for cellulose synthase and family 48 for callose synthase (Cantarel et al, 2009).…”

mentioning

confidence: 99%

Activation of β-Glucan Synthases by Wall-Bound Purple Acid Phosphatase in Tobacco Cells

et al. 2009

View full text Add to dashboard Cite

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 b-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 wildtype 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.

show abstract

“…Callose is also the main component of callose plugs, which form regularly in the mature region and allow cytoplasm to concentrate in the tube apex (Cresti and VanWent, 1976). Callose is produced by the enzyme callose synthase (Brownfield et al, 2007), which is localized in the plasma membrane (Ferguson et al, 1998). Cellulose occurs in lower quantities than callose (Schlupmann et al, 1994) and is generally localized in the inner layer of the cell wall (Ferguson et al, 1998).…”

mentioning

confidence: 99%

Sucrose Synthase Is Associated with the Cell Wall of Tobacco Pollen Tubes

et al. 2008

View full text Add to dashboard Cite

Sucrose synthase (Sus; EC 2.4.1.13) is a key enzyme of sucrose metabolism in plant cells, providing carbon for respiration and for the synthesis of cell wall polymers and starch. Since Sus is important for plant cell growth, insights into its structure, localization, and features are useful for defining the relationships between nutrients, growth, and cell morphogenesis. We used the pollen tube of tobacco (Nicotiana tabacum) as a cell model to characterize the main features of Sus with regard to cell growth and cell wall synthesis. Apart from its role during sexual reproduction, the pollen tube is a typical tip-growing cell, and the proper construction of its cell wall is essential for correct shaping and direction of growth. The outer cell wall layer of pollen tubes consists of pectins, but the inner layer is composed of cellulose and callose; both polymers require metabolic precursors in the form of UDP-glucose, which is synthesized by Sus. We identified an 88-kD polypeptide in the soluble, plasma membrane and Golgi fraction of pollen tubes. The protein was also found in association with the cell wall. After purification, the protein showed an enzyme activity similar to that of maize (Zea mays) Sus. Distribution of Sus was affected by brefeldin A and depended on the nutrition status of the pollen tube, because an absence of metabolic sugars in the growth medium caused Sus to distribute differently during tube elongation. Analysis by bidimensional electrophoresis indicated that Sus exists as two isoforms, one of which is phosphorylated and more abundant in the cytoplasm and cell wall and the other of which is not phosphorylated and is specific to the plasma membrane. Results indicate that the protein has a role in the construction of the extracellular matrix and thus in the morphogenesis of pollen tubes.

show abstract

Proteomic and biochemical evidence links the callose synthase in Nicotiana alata pollen tubes to the product of the NaGSL1 gene

Cited by 58 publications

References 46 publications

A barley cellulose synthase-like CSLH gene mediates (1,3;1,4)-β- d -glucan synthesis in transgenic Arabidopsis

A barley cellulose synthase-like CSLH gene mediates (1,3;1,4)-β- d -glucan synthesis in transgenic Arabidopsis

Activation of β-Glucan Synthases by Wall-Bound Purple Acid Phosphatase in Tobacco Cells

Sucrose Synthase Is Associated with the Cell Wall of Tobacco Pollen Tubes

Contact Info

Product

Resources

About