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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 β-1,4-galactan. Plants in which all three genes were inactivated had no detectable β-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.

Authors:
ORCiD logo [1];  [2];  [2];  [2];  [2];  [3];  [2];  [4];  [2];  [5];  [2];  [6];  [5];  [2];  [5];  [5];  [6];  [7]; ORCiD logo [8]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of Melbourne (Australia); Univ. of Copenhagen (Denmark)
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  3. Univ. of Copenhagen (Denmark)
  4. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of Melbourne (Australia)
  5. Univ. of Melbourne (Australia)
  6. Technical Univ. of Denmark, Kgs. Lyngby (Denmark)
  7. Univ. of Copenhagen, Frederiksberg (Denmark)
  8. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1477373
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Plant and Cell Physiology
Additional Journal Information:
Journal Volume: 59; Journal Issue: 12; Journal ID: ISSN 0032-0781
Publisher:
Japanese Society of Plant Physiologists
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Ebert, Berit, Birdseye, Devon, Liwanag, April J. M., Laursen, Tomas, Rennie, Emilie A., Guo, Xiaoyuan, Catena, Michela, Rautengarten, Carsten, Stonebloom, Solomon H., Gluza, Pawel, Pidatala, Venkataramana R., Andersen, Mathias C. F., Cheetamun, Roshan, Mortimer, Jenny C., Heazlewood, Joshua L., Bacic, Antony, Clausen, Mads H., Willats, William G. T., and Scheller, Henrik V. The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases. United States: N. p., 2018. Web. doi:10.1093/pcp/pcy180.
Ebert, Berit, Birdseye, Devon, Liwanag, April J. M., Laursen, Tomas, Rennie, Emilie A., Guo, Xiaoyuan, Catena, Michela, Rautengarten, Carsten, Stonebloom, Solomon H., Gluza, Pawel, Pidatala, Venkataramana R., Andersen, Mathias C. F., Cheetamun, Roshan, Mortimer, Jenny C., Heazlewood, Joshua L., Bacic, Antony, Clausen, Mads H., Willats, William G. T., & Scheller, Henrik V. The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases. United States. doi:10.1093/pcp/pcy180.
Ebert, Berit, Birdseye, Devon, Liwanag, April J. M., Laursen, Tomas, Rennie, Emilie A., Guo, Xiaoyuan, Catena, Michela, Rautengarten, Carsten, Stonebloom, Solomon H., Gluza, Pawel, Pidatala, Venkataramana R., Andersen, Mathias C. F., Cheetamun, Roshan, Mortimer, Jenny C., Heazlewood, Joshua L., Bacic, Antony, Clausen, Mads H., Willats, William G. T., and Scheller, Henrik V. Sat . "The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases". United States. doi:10.1093/pcp/pcy180. https://www.osti.gov/servlets/purl/1477373.
@article{osti_1477373,
title = {The Three Members of the Arabidopsis Glycosyltransferase Family 92 are Functional β-1,4-Galactan Synthases},
author = {Ebert, Berit and Birdseye, Devon and Liwanag, April J. M. and Laursen, Tomas and Rennie, Emilie A. and Guo, Xiaoyuan and Catena, Michela and Rautengarten, Carsten and Stonebloom, Solomon H. and Gluza, Pawel and Pidatala, Venkataramana R. and Andersen, Mathias C. F. and Cheetamun, Roshan and Mortimer, Jenny C. and Heazlewood, Joshua L. and Bacic, Antony and Clausen, Mads H. and Willats, William G. T. and Scheller, Henrik V.},
abstractNote = {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 β-1,4-galactan. Plants in which all three genes were inactivated had no detectable β-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.},
doi = {10.1093/pcp/pcy180},
journal = {Plant and Cell Physiology},
issn = {0032-0781},
number = 12,
volume = 59,
place = {United States},
year = {2018},
month = {9}
}

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