The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases

Berit Ebert; Devon Birdseye; April J M Liwanag; Tomas Laursen; Emilie A Rennie; Xiaoyuan Guo; Michela Catena; Carsten Rautengarten; Solomon H. Stonebloom; Pawel Gluza; Venkataramana Pidatala; Mathias C F Andersen; Roshan Cheetamun; Jenny C Mortimer; Joshua L Heazlewood; Antony Bacic; Mads H Clausen; William G T Willats; Henrik V. Scheller

doi:10.1093/pcp/pcy180

The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases

Berit Ebert, Devon Birdseye, April J M Liwanag, Tomas Laursen, Emilie A Rennie, Xiaoyuan Guo, Michela Catena, Carsten Rautengarten, Solomon H. Stonebloom, Pawel Gluza, Venkataramana Pidatala, Mathias C F Andersen, Roshan Cheetamun, Jenny C Mortimer, Joshua L Heazlewood, Antony Bacic, Mads H Clausen, William G T Willats, Henrik V. Scheller

12 Citationer (Scopus)

Abstract

Pectin is a major component of primary cell walls and performs a plethora of functions crucial for plant growth, development and plant-defense responses. Despite the importance of pectic polysaccharides their biosynthesis is poorly understood. Several genes have been implicated in pectin biosynthesis by mutant analysis, but biochemical activity has been shown for very few. We used reverse genetics and biochemical analysis to study members of Glycosyltransferase Family 92 (GT92) in Arabidopsis thaliana. Biochemical analysis gave detailed insight into the properties of GALS1 (Galactan synthase 1) and showed galactan synthase activity of GALS2 and GALS3. All proteins are responsible for adding galactose onto existing galactose residues attached to the rhamnogalacturonan-I (RG-I) backbone. Significant GALS activity was observed with galactopentaose as acceptor but longer acceptors are favored. Overexpression of the GALS proteins in Arabidopsis resulted in accumulation of unbranched b-1, 4-galactan. Plants in which all three genes were inactivated had no detectable b-1, 4-galactan, and surprisingly these plants exhibited no obvious developmental phenotypes under standard growth conditions. RG-I in the triple mutants retained branching indicating that the initial Gal substitutions on the RG-I backbone are added by enzymes different from GALS.

Originalsprog	Engelsk
Tidsskrift	Plant and Cell Physiology
Vol/bind	59
Udgave nummer	12
Sider (fra-til)	2624-2636
Antal sider	13
ISSN	0032-0781
DOI	https://doi.org/10.1093/pcp/pcy180
Status	Udgivet - 2018

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10.1093/pcp/pcy180

https://watermark.silverchair.com/pcy180.pdf?token=AQECAHi208BE49Ooan9kkhW_Ercy7Dm3ZL_9Cf3qfKAc485ysgAAAp8wggKbBgkqhkiG9w0BBwagggKMMIICiAIBADCCAoEGCSqGSIb3DQEHATAeBglghkgBZQMEAS4wEQQMOy-_RK_-F6fx1YkTAgEQgIICUl_nC_zIikLgvD1iCpsTswmlrROIBe2CRFSfDlqjgvPp4L8QSPfq7NDES6mmzyHhxtxufpvn4rr9dFT9bM8R6mzN0ExF1mnxb3beAqQnv5XBi_WYVSJn607YwpE1lypR2s-RTgbd6bee1NGNbtIXpwBVNTFH-if3uXmuwnp6brF0tcmeN8KlPIpKELArnjog0KnVHdakVTvtrpYa8ugmIwyZc8fJS9UJUMK0sS2vDLqAEHRNtaq1dHaYcDMeqQ69Q1GgqaJuMFmJ1b-vhnaQI2v0pFxLqB3XCyBuz780TRtSmltclhAYOIcK-bP7bszZNSQf5gKi8R-J3EUcou7pF6wRdF0ILDEy4berwxoL7x8ca0wtGwpp-ZClojBymmVqQ069di19o4GHDC2Yf6RWV_FgulRcndWXe9EZQc_nBj0xCz9wyHyJmFowGUI9LdtqCcL3RgmUV7h4JRzPWk66CL-o7xIxoNS4qCVOmgVenMR0oR2kqqtK4h2CpQOGRB8qNnoVYzFD2qnVM9_hzDfgmPRnZ8WEEcJDLv4-tBNMXizBPW109EFdC8xM38IXNH2tesS_d2KA_H6ArbVEwQNzTlHXQgtbOV4TbJrDthsrLZ5Z4MiWDH2FOdVmM16hpBR1FibWN6aHRbzet3qu0S_B5AzvcskLM5YP-C-9BnKNzMtDdh2Z607HUo9tuSDq_BZwHK_qhdm7FjoGkENyaTM6JX6wdIJny7DNzIYvvmET5v6pg1cVVbnBJizNmP9ghyGw2xOK52_LENjoLdm-DvtZUfNFMw

Citationsformater

Ebert, B., Birdseye, D., Liwanag, A. J. M., Laursen, T., Rennie, E. A., Guo, X., Catena, M., Rautengarten, C., Stonebloom, S. H., Gluza, P., Pidatala, V., Andersen, M. C. F., Cheetamun, R., Mortimer, J. C., Heazlewood, J. L., Bacic, A., Clausen, M. H., Willats, W. G. T., & Scheller, H. V. (2018). The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases. Plant and Cell Physiology, 59(12), 2624-2636. https://doi.org/10.1093/pcp/pcy180

Ebert, B, Birdseye, D, Liwanag, AJM, Laursen, T, Rennie, EA, Guo, X, Catena, M, Rautengarten, C, Stonebloom, SH, Gluza, P, Pidatala, V, Andersen, MCF, Cheetamun, R, Mortimer, JC, Heazlewood, JL, Bacic, A, Clausen, MH, Willats, WGT & Scheller, HV 2018, 'The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases', Plant and Cell Physiology, bind 59, nr. 12, s. 2624-2636. https://doi.org/10.1093/pcp/pcy180

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title = "The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases",

abstract = "Pectin is a major component of primary cell walls and performs a plethora of functions crucial for plant growth, development and plant-defense responses. Despite the importance of pectic polysaccharides their biosynthesis is poorly understood. Several genes have been implicated in pectin biosynthesis by mutant analysis, but biochemical activity has been shown for very few. We used reverse genetics and biochemical analysis to study members of Glycosyltransferase Family 92 (GT92) in Arabidopsis thaliana. Biochemical analysis gave detailed insight into the properties of GALS1 (Galactan synthase 1) and showed galactan synthase activity of GALS2 and GALS3. All proteins are responsible for adding galactose onto existing galactose residues attached to the rhamnogalacturonan-I (RG-I) backbone. Significant GALS activity was observed with galactopentaose as acceptor but longer acceptors are favored. Overexpression of the GALS proteins in Arabidopsis resulted in accumulation of unbranched b-1, 4-galactan. Plants in which all three genes were inactivated had no detectable b-1, 4-galactan, and surprisingly these plants exhibited no obvious developmental phenotypes under standard growth conditions. RG-I in the triple mutants retained branching indicating that the initial Gal substitutions on the RG-I backbone are added by enzymes different from GALS.",

author = "Berit Ebert and Devon Birdseye and Liwanag, {April J M} and Tomas Laursen and Rennie, {Emilie A} and Xiaoyuan Guo and Michela Catena and Carsten Rautengarten and Stonebloom, {Solomon H.} and Pawel Gluza and Venkataramana Pidatala and Andersen, {Mathias C F} and Roshan Cheetamun and Mortimer, {Jenny C} and Heazlewood, {Joshua L} and Antony Bacic and Clausen, {Mads H} and Willats, {William G T} and Scheller, {Henrik V.}",

year = "2018",

doi = "10.1093/pcp/pcy180",

language = "English",

volume = "59",

pages = "2624--2636",

journal = "Plant and Cell Physiology",

issn = "0032-0781",

publisher = "Oxford University Press",

number = "12",

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TY - JOUR

T1 - The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases

AU - Ebert, Berit

AU - Birdseye, Devon

AU - Liwanag, April J M

AU - Laursen, Tomas

AU - Rennie, Emilie A

AU - Guo, Xiaoyuan

AU - Catena, Michela

AU - Rautengarten, Carsten

AU - Stonebloom, Solomon H.

AU - Gluza, Pawel

AU - Pidatala, Venkataramana

AU - Andersen, Mathias C F

AU - Cheetamun, Roshan

AU - Mortimer, Jenny C

AU - Heazlewood, Joshua L

AU - Bacic, Antony

AU - Clausen, Mads H

AU - Willats, William G T

AU - Scheller, Henrik V.

PY - 2018

Y1 - 2018

N2 - Pectin is a major component of primary cell walls and performs a plethora of functions crucial for plant growth, development and plant-defense responses. Despite the importance of pectic polysaccharides their biosynthesis is poorly understood. Several genes have been implicated in pectin biosynthesis by mutant analysis, but biochemical activity has been shown for very few. We used reverse genetics and biochemical analysis to study members of Glycosyltransferase Family 92 (GT92) in Arabidopsis thaliana. Biochemical analysis gave detailed insight into the properties of GALS1 (Galactan synthase 1) and showed galactan synthase activity of GALS2 and GALS3. All proteins are responsible for adding galactose onto existing galactose residues attached to the rhamnogalacturonan-I (RG-I) backbone. Significant GALS activity was observed with galactopentaose as acceptor but longer acceptors are favored. Overexpression of the GALS proteins in Arabidopsis resulted in accumulation of unbranched b-1, 4-galactan. Plants in which all three genes were inactivated had no detectable b-1, 4-galactan, and surprisingly these plants exhibited no obvious developmental phenotypes under standard growth conditions. RG-I in the triple mutants retained branching indicating that the initial Gal substitutions on the RG-I backbone are added by enzymes different from GALS.

AB - Pectin is a major component of primary cell walls and performs a plethora of functions crucial for plant growth, development and plant-defense responses. Despite the importance of pectic polysaccharides their biosynthesis is poorly understood. Several genes have been implicated in pectin biosynthesis by mutant analysis, but biochemical activity has been shown for very few. We used reverse genetics and biochemical analysis to study members of Glycosyltransferase Family 92 (GT92) in Arabidopsis thaliana. Biochemical analysis gave detailed insight into the properties of GALS1 (Galactan synthase 1) and showed galactan synthase activity of GALS2 and GALS3. All proteins are responsible for adding galactose onto existing galactose residues attached to the rhamnogalacturonan-I (RG-I) backbone. Significant GALS activity was observed with galactopentaose as acceptor but longer acceptors are favored. Overexpression of the GALS proteins in Arabidopsis resulted in accumulation of unbranched b-1, 4-galactan. Plants in which all three genes were inactivated had no detectable b-1, 4-galactan, and surprisingly these plants exhibited no obvious developmental phenotypes under standard growth conditions. RG-I in the triple mutants retained branching indicating that the initial Gal substitutions on the RG-I backbone are added by enzymes different from GALS.

U2 - 10.1093/pcp/pcy180

DO - 10.1093/pcp/pcy180

M3 - Journal article

C2 - 30184190

SN - 0032-0781

VL - 59

SP - 2624

EP - 2636

JO - Plant and Cell Physiology

JF - Plant and Cell Physiology

IS - 12

ER -

The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases

Abstract

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