Arabidopsis thaliana alpha1,2-glucosyltransferase (ALG10) is required for efficient N-glycosylation and leaf growth

Plant J. 2011 Oct;68(2):314-25. doi: 10.1111/j.1365-313X.2011.04688.x. Epub 2011 Jul 27.

Abstract

Assembly of the dolichol-linked oligosaccharide precursor (Glc(3) Man(9) GlcNAc(2) ) is highly conserved among eukaryotes. In contrast to yeast and mammals, little is known about the biosynthesis of dolichol-linked oligosaccharides and the transfer to asparagine residues of nascent polypeptides in plants. To understand the biological function of these processes in plants we characterized the Arabidopsis thaliana homolog of yeast ALG10, the α1,2-glucosyltransferase that transfers the terminal glucose residue to the lipid-linked precursor. Expression of an Arabidopsis ALG10-GFP fusion protein in Nicotiana benthamiana leaf epidermal cells revealed a reticular distribution pattern resembling endoplasmic reticulum (ER) localization. Analysis of lipid-linked oligosaccharides showed that Arabidopsis ALG10 can complement the yeast Δalg10 mutant strain. A homozygous Arabidopsis T-DNA insertion mutant (alg10-1) accumulated mainly lipid-linked Glc(2) Man(9) GlcNAc(2) and displayed a severe protein underglycosylation defect. Phenotypic analysis of alg10-1 showed that mutant plants have altered leaf size when grown in soil. Moreover, the inactivation of ALG10 in Arabidopsis resulted in the activation of the unfolded protein response, increased salt sensitivity and suppression of the phenotype of α-glucosidase I-deficient plants. In summary, these data show that Arabidopsis ALG10 is an ER-resident α1,2-glucosyltransferase that is required for lipid-linked oligosaccharide biosynthesis and subsequently for normal leaf development and abiotic stress response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / embryology
  • Arabidopsis / enzymology*
  • Arabidopsis / genetics
  • Arabidopsis / growth & development
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Carbohydrate Sequence
  • Endoplasmic Reticulum / enzymology
  • Endoplasmic Reticulum / metabolism
  • Genetic Complementation Test
  • Glucosyltransferases / genetics
  • Glucosyltransferases / metabolism
  • Glycosylation
  • Green Fluorescent Proteins
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Oligosaccharides / biosynthesis*
  • Oligosaccharides / chemistry
  • Phenotype
  • Plant Leaves / enzymology
  • Plant Leaves / genetics
  • Plant Leaves / growth & development*
  • Plant Leaves / metabolism
  • Polyisoprenyl Phosphate Sugars / biosynthesis*
  • Polyisoprenyl Phosphate Sugars / chemistry
  • Polysaccharides / metabolism
  • Protein Processing, Post-Translational
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Salt Tolerance
  • Stress, Physiological
  • Tobacco / genetics
  • Tobacco / metabolism
  • Unfolded Protein Response

Substances

  • Arabidopsis Proteins
  • Glc(3)Man(9)(GlcNAc)(2)-diphosphate-dolichol
  • Oligosaccharides
  • Polyisoprenyl Phosphate Sugars
  • Polysaccharides
  • Green Fluorescent Proteins
  • ALG10 protein, Arabidopsis
  • Glucosyltransferases