A β-glucuronosyltransferase from Arabidopsis thaliana involved in biosynthesis of type II arabinogalactan has a role in cell elongation during seedling growth

Eva Knoch; Adiphol Dilokpimol; Theodora Tryfona; Christian Peter Poulsen; Guangyan Xiong; Jesper Harholt; Bent L Petersen; Peter Ulvskov; Masood Z. Hadi; Toshihisa Kotake; Yoichi Tsumuraya; Markus Pauly; Paul Dupree; Naomi Geshi

doi:10.1111/tpj.12353

A β-glucuronosyltransferase from Arabidopsis thaliana involved in biosynthesis of type II arabinogalactan has a role in cell elongation during seedling growth

Eva Knoch, Adiphol Dilokpimol, Theodora Tryfona, Christian Peter Poulsen, Guangyan Xiong, Jesper Harholt, Bent L Petersen, Peter Ulvskov, Masood Z. Hadi, Toshihisa Kotake, Yoichi Tsumuraya, Markus Pauly, Paul Dupree, Naomi Geshi

58 Citations (Scopus)

Abstract

We have characterized a β-glucuronosyltransferase (AtGlcAT14A) from Arabidopsis thaliana that is involved in the biosynthesis of type II arabinogalactan (AG). This enzyme belongs to the Carbohydrate Active Enzyme database glycosyltransferase family 14 (GT14). The protein was localized to the Golgi apparatus when transiently expressed in Nicotiana benthamiana. The soluble catalytic domain expressed in Pichia pastoris transferred glucuronic acid (GlcA) to β-1,6-galactooligosaccharides with degrees of polymerization (DP) ranging from 3-11, and to β-1,3-galactooligosaccharides of DP5 and 7, indicating that the enzyme is a glucuronosyltransferase that modifies both the β-1,6- and β-1,3-galactan present in type II AG. Two allelic T-DNA insertion mutant lines showed 20-35% enhanced cell elongation during seedling growth compared to wild-type. Analyses of AG isolated from the mutants revealed a reduction of GlcA substitution on Gal-β-1,6-Gal and β-1,3-Gal, indicating an in vivo role of AtGlcAT14A in synthesis of those structures in type II AG. Moreover, a relative increase in the levels of 3-, 6- and 3,6-linked galactose (Gal) and reduced levels of 3-, 2- and 2,5-linked arabinose (Ara) were seen, suggesting that the mutation in AtGlcAT14A results in a relative increase of the longer and branched β-1,3- and β-1,6-galactans. This increase of galactosylation in the mutants is most likely caused by increased availability of the O6 position of Gal, which is a shared acceptor site for AtGlcAT14A and galactosyltransferases in synthesis of type II AG, and thus addition of GlcA may terminate Gal chain extension. We discuss a role for the glucuronosyltransferase in the biosynthesis of type II AG, with a biological role during seedling growth.

Original language	English
Journal	Plant Journal
Volume	76
Issue number	6
Pages (from-to)	1016-1029
Number of pages	14
ISSN	0960-7412
DOIs	https://doi.org/10.1111/tpj.12353
Publication status	Published - Dec 2013

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Knoch, E., Dilokpimol, A., Tryfona, T., Poulsen, C. P., Xiong, G., Harholt, J., Petersen, B. L., Ulvskov, P., Hadi, M. Z., Kotake, T., Tsumuraya, Y., Pauly, M., Dupree, P., & Geshi, N. (2013). A β-glucuronosyltransferase from Arabidopsis thaliana involved in biosynthesis of type II arabinogalactan has a role in cell elongation during seedling growth. Plant Journal, 76(6), 1016-1029. https://doi.org/10.1111/tpj.12353

Knoch, E, Dilokpimol, A, Tryfona, T, Poulsen, CP, Xiong, G, Harholt, J, Petersen, BL , Ulvskov, P, Hadi, MZ, Kotake, T, Tsumuraya, Y, Pauly, M, Dupree, P & Geshi, N 2013, 'A β-glucuronosyltransferase from Arabidopsis thaliana involved in biosynthesis of type II arabinogalactan has a role in cell elongation during seedling growth', Plant Journal, vol. 76, no. 6, pp. 1016-1029. https://doi.org/10.1111/tpj.12353

@article{69bf6be2a5804c03896f23b82cd36a02,

title = "A β-glucuronosyltransferase from Arabidopsis thaliana involved in biosynthesis of type II arabinogalactan has a role in cell elongation during seedling growth",

abstract = "We have characterized a β-glucuronosyltransferase (AtGlcAT14A) from Arabidopsis thaliana that is involved in the biosynthesis of type II arabinogalactan (AG). This enzyme belongs to the Carbohydrate Active Enzyme database glycosyltransferase family 14 (GT14). The protein was localized to the Golgi apparatus when transiently expressed in Nicotiana benthamiana. The soluble catalytic domain expressed in Pichia pastoris transferred glucuronic acid (GlcA) to β-1,6-galactooligosaccharides with degrees of polymerization (DP) ranging from 3-11, and to β-1,3-galactooligosaccharides of DP5 and 7, indicating that the enzyme is a glucuronosyltransferase that modifies both the β-1,6- and β-1,3-galactan present in type II AG. Two allelic T-DNA insertion mutant lines showed 20-35% enhanced cell elongation during seedling growth compared to wild-type. Analyses of AG isolated from the mutants revealed a reduction of GlcA substitution on Gal-β-1,6-Gal and β-1,3-Gal, indicating an in vivo role of AtGlcAT14A in synthesis of those structures in type II AG. Moreover, a relative increase in the levels of 3-, 6- and 3,6-linked galactose (Gal) and reduced levels of 3-, 2- and 2,5-linked arabinose (Ara) were seen, suggesting that the mutation in AtGlcAT14A results in a relative increase of the longer and branched β-1,3- and β-1,6-galactans. This increase of galactosylation in the mutants is most likely caused by increased availability of the O6 position of Gal, which is a shared acceptor site for AtGlcAT14A and galactosyltransferases in synthesis of type II AG, and thus addition of GlcA may terminate Gal chain extension. We discuss a role for the glucuronosyltransferase in the biosynthesis of type II AG, with a biological role during seedling growth.",

author = "Eva Knoch and Adiphol Dilokpimol and Theodora Tryfona and Poulsen, {Christian Peter} and Guangyan Xiong and Jesper Harholt and Petersen, {Bent L} and Peter Ulvskov and Hadi, {Masood Z.} and Toshihisa Kotake and Yoichi Tsumuraya and Markus Pauly and Paul Dupree and Naomi Geshi",

year = "2013",

month = dec,

doi = "10.1111/tpj.12353",

language = "English",

volume = "76",

pages = "1016--1029",

journal = "Plant Journal",

issn = "0960-7412",

publisher = "Wiley-Blackwell",

number = "6",

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TY - JOUR

T1 - A β-glucuronosyltransferase from Arabidopsis thaliana involved in biosynthesis of type II arabinogalactan has a role in cell elongation during seedling growth

AU - Knoch, Eva

AU - Dilokpimol, Adiphol

AU - Tryfona, Theodora

AU - Poulsen, Christian Peter

AU - Xiong, Guangyan

AU - Harholt, Jesper

AU - Petersen, Bent L

AU - Ulvskov, Peter

AU - Hadi, Masood Z.

AU - Kotake, Toshihisa

AU - Tsumuraya, Yoichi

AU - Pauly, Markus

AU - Dupree, Paul

AU - Geshi, Naomi

PY - 2013/12

Y1 - 2013/12

N2 - We have characterized a β-glucuronosyltransferase (AtGlcAT14A) from Arabidopsis thaliana that is involved in the biosynthesis of type II arabinogalactan (AG). This enzyme belongs to the Carbohydrate Active Enzyme database glycosyltransferase family 14 (GT14). The protein was localized to the Golgi apparatus when transiently expressed in Nicotiana benthamiana. The soluble catalytic domain expressed in Pichia pastoris transferred glucuronic acid (GlcA) to β-1,6-galactooligosaccharides with degrees of polymerization (DP) ranging from 3-11, and to β-1,3-galactooligosaccharides of DP5 and 7, indicating that the enzyme is a glucuronosyltransferase that modifies both the β-1,6- and β-1,3-galactan present in type II AG. Two allelic T-DNA insertion mutant lines showed 20-35% enhanced cell elongation during seedling growth compared to wild-type. Analyses of AG isolated from the mutants revealed a reduction of GlcA substitution on Gal-β-1,6-Gal and β-1,3-Gal, indicating an in vivo role of AtGlcAT14A in synthesis of those structures in type II AG. Moreover, a relative increase in the levels of 3-, 6- and 3,6-linked galactose (Gal) and reduced levels of 3-, 2- and 2,5-linked arabinose (Ara) were seen, suggesting that the mutation in AtGlcAT14A results in a relative increase of the longer and branched β-1,3- and β-1,6-galactans. This increase of galactosylation in the mutants is most likely caused by increased availability of the O6 position of Gal, which is a shared acceptor site for AtGlcAT14A and galactosyltransferases in synthesis of type II AG, and thus addition of GlcA may terminate Gal chain extension. We discuss a role for the glucuronosyltransferase in the biosynthesis of type II AG, with a biological role during seedling growth.

AB - We have characterized a β-glucuronosyltransferase (AtGlcAT14A) from Arabidopsis thaliana that is involved in the biosynthesis of type II arabinogalactan (AG). This enzyme belongs to the Carbohydrate Active Enzyme database glycosyltransferase family 14 (GT14). The protein was localized to the Golgi apparatus when transiently expressed in Nicotiana benthamiana. The soluble catalytic domain expressed in Pichia pastoris transferred glucuronic acid (GlcA) to β-1,6-galactooligosaccharides with degrees of polymerization (DP) ranging from 3-11, and to β-1,3-galactooligosaccharides of DP5 and 7, indicating that the enzyme is a glucuronosyltransferase that modifies both the β-1,6- and β-1,3-galactan present in type II AG. Two allelic T-DNA insertion mutant lines showed 20-35% enhanced cell elongation during seedling growth compared to wild-type. Analyses of AG isolated from the mutants revealed a reduction of GlcA substitution on Gal-β-1,6-Gal and β-1,3-Gal, indicating an in vivo role of AtGlcAT14A in synthesis of those structures in type II AG. Moreover, a relative increase in the levels of 3-, 6- and 3,6-linked galactose (Gal) and reduced levels of 3-, 2- and 2,5-linked arabinose (Ara) were seen, suggesting that the mutation in AtGlcAT14A results in a relative increase of the longer and branched β-1,3- and β-1,6-galactans. This increase of galactosylation in the mutants is most likely caused by increased availability of the O6 position of Gal, which is a shared acceptor site for AtGlcAT14A and galactosyltransferases in synthesis of type II AG, and thus addition of GlcA may terminate Gal chain extension. We discuss a role for the glucuronosyltransferase in the biosynthesis of type II AG, with a biological role during seedling growth.

U2 - 10.1111/tpj.12353

DO - 10.1111/tpj.12353

M3 - Journal article

C2 - 24128328

SN - 0960-7412

VL - 76

SP - 1016

EP - 1029

JO - Plant Journal

JF - Plant Journal

IS - 6

ER -

A β-glucuronosyltransferase from Arabidopsis thaliana involved in biosynthesis of type II arabinogalactan has a role in cell elongation during seedling growth

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