Identification of the Gene Encoding the 1,3-Mannosyltransferase (ALG3) in Arabidopsis and Characterization of Downstream N-Glycan Processing[W].

M.G.L. Henquet, L. Lehle, M.E.L. Schreuder, G.J.A. Rouwendal, J.W. Molthoff, J.P.F.G. Helsper, A.R. van der Krol, H.J. Bosch

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Abstract

Glycosyltransferases are involved in the biosynthesis of lipid-linked N-glycans. Here, we identify and characterize a mannosyltransferase gene from Arabidopsis thaliana, which is the functional homolog of the ALG3 (Dol-P-Man: Man(5)GlcNAc(2)-PP-Dol alpha 1,3-mannosyl transferase) gene in yeast. The At ALG3 protein can complement a Delta alg3 yeast mutant and is localized to the endoplasmic reticulum in yeast and in plants. A homozygous T-DNA insertion mutant, alg3-2, was identified in Arabidopsis with residual levels of wild-type ALG3, derived from incidental splicing of the 11th intron carrying the T-DNAs. N- glycan analysis of alg3-2 and alg3-2 in the complex-glycan-less mutant background, which lacks N- acetylglucosaminyl-transferase I activity, reveals that when ALG3 activity is strongly reduced, almost all N-glycans transferred to proteins are aberrant, indicating that the Arabidopsis oligosaccharide transferase complex is remarkably substrate tolerant. In alg3-2 plants, the aberrant glycans on glycoproteins are recognized by endogenous mannosidase I and N- acetylglucosaminyltransferase I and efficiently processed into complex-type glycans. Although no high-mannose-type glycoproteins are detected in alg3-2 plants, these plants do not show a growth phenotype under normal growth conditions. However, the glycosylation abnormalities result in activation of marker genes diagnostic of the unfolded protein response.
Original languageEnglish
Pages (from-to)1652-1664
JournalThe Plant Cell
Volume20
Issue number6
DOIs
Publication statusPublished - 2008

Fingerprint

Mannosyltransferases
Arabidopsis
Polysaccharides
polysaccharides
transferases
Genes
genes
Yeasts
mannosyl-oligosaccharide 1,2-alpha-mannosidase
Transferases
yeasts
mutants
glycoproteins
Glycoproteins
mannosidases
unfolded protein response
Unfolded Protein Response
Glycosyltransferases
glycosyltransferases
DNA

Keywords

  • asparagine-linked oligosaccharides
  • deficient glycoprotein syndrome
  • unfolded protein response
  • yeast mutants deficient
  • endoplasmic-reticulum
  • saccharomyces-cerevisiae
  • congenital disorder
  • oligosaccharyltransferase complex
  • acetylglucosaminyltransferase

Cite this

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title = "Identification of the Gene Encoding the 1,3-Mannosyltransferase (ALG3) in Arabidopsis and Characterization of Downstream N-Glycan Processing[W].",
abstract = "Glycosyltransferases are involved in the biosynthesis of lipid-linked N-glycans. Here, we identify and characterize a mannosyltransferase gene from Arabidopsis thaliana, which is the functional homolog of the ALG3 (Dol-P-Man: Man(5)GlcNAc(2)-PP-Dol alpha 1,3-mannosyl transferase) gene in yeast. The At ALG3 protein can complement a Delta alg3 yeast mutant and is localized to the endoplasmic reticulum in yeast and in plants. A homozygous T-DNA insertion mutant, alg3-2, was identified in Arabidopsis with residual levels of wild-type ALG3, derived from incidental splicing of the 11th intron carrying the T-DNAs. N- glycan analysis of alg3-2 and alg3-2 in the complex-glycan-less mutant background, which lacks N- acetylglucosaminyl-transferase I activity, reveals that when ALG3 activity is strongly reduced, almost all N-glycans transferred to proteins are aberrant, indicating that the Arabidopsis oligosaccharide transferase complex is remarkably substrate tolerant. In alg3-2 plants, the aberrant glycans on glycoproteins are recognized by endogenous mannosidase I and N- acetylglucosaminyltransferase I and efficiently processed into complex-type glycans. Although no high-mannose-type glycoproteins are detected in alg3-2 plants, these plants do not show a growth phenotype under normal growth conditions. However, the glycosylation abnormalities result in activation of marker genes diagnostic of the unfolded protein response.",
keywords = "asparagine-linked oligosaccharides, deficient glycoprotein syndrome, unfolded protein response, yeast mutants deficient, endoplasmic-reticulum, saccharomyces-cerevisiae, congenital disorder, oligosaccharyltransferase complex, acetylglucosaminyltransferase",
author = "M.G.L. Henquet and L. Lehle and M.E.L. Schreuder and G.J.A. Rouwendal and J.W. Molthoff and J.P.F.G. Helsper and {van der Krol}, A.R. and H.J. Bosch",
year = "2008",
doi = "10.1105/tpc.108.060731",
language = "English",
volume = "20",
pages = "1652--1664",
journal = "The Plant Cell",
issn = "1040-4651",
publisher = "American Society of Plant Biologists",
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}

Identification of the Gene Encoding the 1,3-Mannosyltransferase (ALG3) in Arabidopsis and Characterization of Downstream N-Glycan Processing[W]. / Henquet, M.G.L.; Lehle, L.; Schreuder, M.E.L.; Rouwendal, G.J.A.; Molthoff, J.W.; Helsper, J.P.F.G.; van der Krol, A.R.; Bosch, H.J.

In: The Plant Cell, Vol. 20, No. 6, 2008, p. 1652-1664.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Identification of the Gene Encoding the 1,3-Mannosyltransferase (ALG3) in Arabidopsis and Characterization of Downstream N-Glycan Processing[W].

AU - Henquet, M.G.L.

AU - Lehle, L.

AU - Schreuder, M.E.L.

AU - Rouwendal, G.J.A.

AU - Molthoff, J.W.

AU - Helsper, J.P.F.G.

AU - van der Krol, A.R.

AU - Bosch, H.J.

PY - 2008

Y1 - 2008

N2 - Glycosyltransferases are involved in the biosynthesis of lipid-linked N-glycans. Here, we identify and characterize a mannosyltransferase gene from Arabidopsis thaliana, which is the functional homolog of the ALG3 (Dol-P-Man: Man(5)GlcNAc(2)-PP-Dol alpha 1,3-mannosyl transferase) gene in yeast. The At ALG3 protein can complement a Delta alg3 yeast mutant and is localized to the endoplasmic reticulum in yeast and in plants. A homozygous T-DNA insertion mutant, alg3-2, was identified in Arabidopsis with residual levels of wild-type ALG3, derived from incidental splicing of the 11th intron carrying the T-DNAs. N- glycan analysis of alg3-2 and alg3-2 in the complex-glycan-less mutant background, which lacks N- acetylglucosaminyl-transferase I activity, reveals that when ALG3 activity is strongly reduced, almost all N-glycans transferred to proteins are aberrant, indicating that the Arabidopsis oligosaccharide transferase complex is remarkably substrate tolerant. In alg3-2 plants, the aberrant glycans on glycoproteins are recognized by endogenous mannosidase I and N- acetylglucosaminyltransferase I and efficiently processed into complex-type glycans. Although no high-mannose-type glycoproteins are detected in alg3-2 plants, these plants do not show a growth phenotype under normal growth conditions. However, the glycosylation abnormalities result in activation of marker genes diagnostic of the unfolded protein response.

AB - Glycosyltransferases are involved in the biosynthesis of lipid-linked N-glycans. Here, we identify and characterize a mannosyltransferase gene from Arabidopsis thaliana, which is the functional homolog of the ALG3 (Dol-P-Man: Man(5)GlcNAc(2)-PP-Dol alpha 1,3-mannosyl transferase) gene in yeast. The At ALG3 protein can complement a Delta alg3 yeast mutant and is localized to the endoplasmic reticulum in yeast and in plants. A homozygous T-DNA insertion mutant, alg3-2, was identified in Arabidopsis with residual levels of wild-type ALG3, derived from incidental splicing of the 11th intron carrying the T-DNAs. N- glycan analysis of alg3-2 and alg3-2 in the complex-glycan-less mutant background, which lacks N- acetylglucosaminyl-transferase I activity, reveals that when ALG3 activity is strongly reduced, almost all N-glycans transferred to proteins are aberrant, indicating that the Arabidopsis oligosaccharide transferase complex is remarkably substrate tolerant. In alg3-2 plants, the aberrant glycans on glycoproteins are recognized by endogenous mannosidase I and N- acetylglucosaminyltransferase I and efficiently processed into complex-type glycans. Although no high-mannose-type glycoproteins are detected in alg3-2 plants, these plants do not show a growth phenotype under normal growth conditions. However, the glycosylation abnormalities result in activation of marker genes diagnostic of the unfolded protein response.

KW - asparagine-linked oligosaccharides

KW - deficient glycoprotein syndrome

KW - unfolded protein response

KW - yeast mutants deficient

KW - endoplasmic-reticulum

KW - saccharomyces-cerevisiae

KW - congenital disorder

KW - oligosaccharyltransferase complex

KW - acetylglucosaminyltransferase

U2 - 10.1105/tpc.108.060731

DO - 10.1105/tpc.108.060731

M3 - Article

VL - 20

SP - 1652

EP - 1664

JO - The Plant Cell

JF - The Plant Cell

SN - 1040-4651

IS - 6

ER -