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Title: Isolation of a mutant Arabidopsis plant that lacks N-aetyl glucosaminyl transferase I and is unable to synthesize Golgi-modified complex N-linked glycans

Abstract

The complex asparagine-linked glycans of plant glycoproteins, characterized by the presence of [beta]1[yields]2 xylose and [alpha]1[yields]3 fucose residues, are derived from typical mannose[sub 9](N-acetylglucosamine)[sub 2] (Man[sub 9]GlcNAc[sub 2]) N-linked glycans through the activity of a series of glycosidases and glycosyl transferases in the Golgi apparatus. By screening leaf extracts with an antiserum against complex glycans, we isolated a mutant of Arbidopsis thaliana that is blocked in the conversion of high-manne to complex glycans. In callus tissues derived from the mutant plants, all glycans bind to concanavalin A. These glycans can be released by treatment with endoglycosidase H, and the majority has the same size as Man[sub 5]GlcNAc[sub 1] glycans. In the presence of deoxymannojirimycin, an inhibitor of mannosidase I, the mutant cells synthesize Man[sub 9]GlcNAc[sub 2] and Man[sub 8]GlcNAc[sub 2] glycans, suggesting that the biochemical lesion in the mutant is not in the biosynthesis of high-mannose glycans in the endoplasmic reticulum but in their modification in the Golgi. Direct enzyme assays of cell extracts show that the mutant cells lack N-acetyl glucosaminyl transferase I, the first enzyme in the pathway of complex glycan biosynthesis. The mutant plants are able to complete their development normally under several environmental conditions, suggesting thatmore » complex glycans are not essential for normal developmental processes. By crossing the complex-glycan-deficient strain of A. thaliana with a transgenic strain that expresses the glycoprotein phytohemagglutinin, a unique strain was obtained that synthesizes phytohemagglutinin with two high-mannose glycans, instead of one high-mannose and one complex glycan. 42 refs., 8 figs., 1 tab.« less

Authors:
; ;  [1];  [2]
  1. Univ. of California, San Diego, La Jolla, CA (United States)
  2. Friedrich Miescher Institute, Basel (Switzerland)
Publication Date:
OSTI Identifier:
5815233
Resource Type:
Journal Article
Journal Name:
Plant Physiology; (United States)
Additional Journal Information:
Journal Volume: 102:4; Journal ID: ISSN 0032-0889
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ARABIDOPSIS; PLANT GROWTH; GLYCOPROTEINS; BIOCHEMISTRY; PHYTOHEMAGGLUTININ; BIOSYNTHESIS; ASPARAGINE; CONCANAVALIN A; ENDOPLASMIC RETICULUM; GLYCOSYL TRANSFERASES; MANNOSE; PLANTS; XYLOSE; AGGLUTININS; ALDEHYDES; AMIDES; AMINO ACIDS; ANTIBODIES; CARBOHYDRATES; CARBOXYLIC ACIDS; CELL CONSTITUENTS; CHEMISTRY; ENZYMES; GROWTH; HEMAGGLUTININS; HEXOSES; LECTINS; MAGNOLIOPHYTA; MAGNOLIOPSIDA; MITOGENS; MONOSACCHARIDES; MUCOPROTEINS; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PENTOSES; POLYSACCHARIDES; PROTEINS; SACCHARIDES; SYNTHESIS; TRANSFERASES; 550200* - Biochemistry

Citation Formats

Schaewen, A von, O'Neill, J, Chrispeels, M J, and Sturm, A. Isolation of a mutant Arabidopsis plant that lacks N-aetyl glucosaminyl transferase I and is unable to synthesize Golgi-modified complex N-linked glycans. United States: N. p., 1993. Web. doi:10.1104/pp.102.4.1109.
Schaewen, A von, O'Neill, J, Chrispeels, M J, & Sturm, A. Isolation of a mutant Arabidopsis plant that lacks N-aetyl glucosaminyl transferase I and is unable to synthesize Golgi-modified complex N-linked glycans. United States. doi:10.1104/pp.102.4.1109.
Schaewen, A von, O'Neill, J, Chrispeels, M J, and Sturm, A. Sun . "Isolation of a mutant Arabidopsis plant that lacks N-aetyl glucosaminyl transferase I and is unable to synthesize Golgi-modified complex N-linked glycans". United States. doi:10.1104/pp.102.4.1109.
@article{osti_5815233,
title = {Isolation of a mutant Arabidopsis plant that lacks N-aetyl glucosaminyl transferase I and is unable to synthesize Golgi-modified complex N-linked glycans},
author = {Schaewen, A von and O'Neill, J and Chrispeels, M J and Sturm, A},
abstractNote = {The complex asparagine-linked glycans of plant glycoproteins, characterized by the presence of [beta]1[yields]2 xylose and [alpha]1[yields]3 fucose residues, are derived from typical mannose[sub 9](N-acetylglucosamine)[sub 2] (Man[sub 9]GlcNAc[sub 2]) N-linked glycans through the activity of a series of glycosidases and glycosyl transferases in the Golgi apparatus. By screening leaf extracts with an antiserum against complex glycans, we isolated a mutant of Arbidopsis thaliana that is blocked in the conversion of high-manne to complex glycans. In callus tissues derived from the mutant plants, all glycans bind to concanavalin A. These glycans can be released by treatment with endoglycosidase H, and the majority has the same size as Man[sub 5]GlcNAc[sub 1] glycans. In the presence of deoxymannojirimycin, an inhibitor of mannosidase I, the mutant cells synthesize Man[sub 9]GlcNAc[sub 2] and Man[sub 8]GlcNAc[sub 2] glycans, suggesting that the biochemical lesion in the mutant is not in the biosynthesis of high-mannose glycans in the endoplasmic reticulum but in their modification in the Golgi. Direct enzyme assays of cell extracts show that the mutant cells lack N-acetyl glucosaminyl transferase I, the first enzyme in the pathway of complex glycan biosynthesis. The mutant plants are able to complete their development normally under several environmental conditions, suggesting that complex glycans are not essential for normal developmental processes. By crossing the complex-glycan-deficient strain of A. thaliana with a transgenic strain that expresses the glycoprotein phytohemagglutinin, a unique strain was obtained that synthesizes phytohemagglutinin with two high-mannose glycans, instead of one high-mannose and one complex glycan. 42 refs., 8 figs., 1 tab.},
doi = {10.1104/pp.102.4.1109},
journal = {Plant Physiology; (United States)},
issn = {0032-0889},
number = ,
volume = 102:4,
place = {United States},
year = {1993},
month = {8}
}