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Title: A Golgi UDP-GlcNAc transporter delivers substrates for N-linked glycans and sphingolipids

Abstract

Glycosylation requires activated glycosyl donors in the form of nucleotide sugars to drive processes such as post-translational protein modifications and glycolipid and polysaccharide biosynthesis. Most of these reactions occur in the Golgi, requiring cytosolic-derived nucleotide sugars, which need to be actively transferred into the Golgi lumen by nucleotide sugar transporters. We identified a Golgi-localized nucleotide sugar transporter from Arabidopsis thaliana with affinity for UDP-N-acetyl-d-glucosamine (UDP-GlcNAc) and assigned it UDP-GlcNAc transporter 1 (UGNT1). Profiles of N-glycopeptides revealed that plants carrying the ugnt1 loss-of-function allele are virtually devoid of complex and hybrid N-glycans. Instead, the N-glycopeptide population from these alleles exhibited high-mannose structures, representing structures prior to the addition of the first GlcNAc in the Golgi. Concomitantly, sphingolipid profiling revealed that the biosynthesis of GlcNAc-containing glycosyl inositol phosphorylceramides (GIPCs) is also reliant on this transporter. By contrast, plants carrying the loss-of-function alleles affecting ROCK1, which has been reported to transport UDP-GlcNAc and UDP-N-acetylgalactosamine, exhibit no changes in N-glycan or GIPC profiles. Our findings reveal that plants contain a single UDP-GlcNAc transporter that delivers an essential substrate for the maturation of N-glycans and the GIPC class of sphingolipids.

Authors:
; ; ; ; ; ; ; ; ; ; ORCiD logo
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:
1478344
DOE Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article
Journal Name:
Nature Plants
Additional Journal Information:
Journal Volume: 4; Journal Issue: 10; Journal ID: ISSN 2055-0278
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Ebert, Berit, Rautengarten, Carsten, McFarlane, Heather E., Rupasinghe, Thusitha, Zeng, Wei, Ford, Kristina, Scheller, Henrik V., Bacic, Antony, Roessner, Ute, Persson, Staffan, and Heazlewood, Joshua L. A Golgi UDP-GlcNAc transporter delivers substrates for N-linked glycans and sphingolipids. United States: N. p., 2018. Web. doi:10.1038/s41477-018-0235-5.
Ebert, Berit, Rautengarten, Carsten, McFarlane, Heather E., Rupasinghe, Thusitha, Zeng, Wei, Ford, Kristina, Scheller, Henrik V., Bacic, Antony, Roessner, Ute, Persson, Staffan, & Heazlewood, Joshua L. A Golgi UDP-GlcNAc transporter delivers substrates for N-linked glycans and sphingolipids. United States. doi:10.1038/s41477-018-0235-5.
Ebert, Berit, Rautengarten, Carsten, McFarlane, Heather E., Rupasinghe, Thusitha, Zeng, Wei, Ford, Kristina, Scheller, Henrik V., Bacic, Antony, Roessner, Ute, Persson, Staffan, and Heazlewood, Joshua L. Mon . "A Golgi UDP-GlcNAc transporter delivers substrates for N-linked glycans and sphingolipids". United States. doi:10.1038/s41477-018-0235-5.
@article{osti_1478344,
title = {A Golgi UDP-GlcNAc transporter delivers substrates for N-linked glycans and sphingolipids},
author = {Ebert, Berit and Rautengarten, Carsten and McFarlane, Heather E. and Rupasinghe, Thusitha and Zeng, Wei and Ford, Kristina and Scheller, Henrik V. and Bacic, Antony and Roessner, Ute and Persson, Staffan and Heazlewood, Joshua L.},
abstractNote = {Glycosylation requires activated glycosyl donors in the form of nucleotide sugars to drive processes such as post-translational protein modifications and glycolipid and polysaccharide biosynthesis. Most of these reactions occur in the Golgi, requiring cytosolic-derived nucleotide sugars, which need to be actively transferred into the Golgi lumen by nucleotide sugar transporters. We identified a Golgi-localized nucleotide sugar transporter from Arabidopsis thaliana with affinity for UDP-N-acetyl-d-glucosamine (UDP-GlcNAc) and assigned it UDP-GlcNAc transporter 1 (UGNT1). Profiles of N-glycopeptides revealed that plants carrying the ugnt1 loss-of-function allele are virtually devoid of complex and hybrid N-glycans. Instead, the N-glycopeptide population from these alleles exhibited high-mannose structures, representing structures prior to the addition of the first GlcNAc in the Golgi. Concomitantly, sphingolipid profiling revealed that the biosynthesis of GlcNAc-containing glycosyl inositol phosphorylceramides (GIPCs) is also reliant on this transporter. By contrast, plants carrying the loss-of-function alleles affecting ROCK1, which has been reported to transport UDP-GlcNAc and UDP-N-acetylgalactosamine, exhibit no changes in N-glycan or GIPC profiles. Our findings reveal that plants contain a single UDP-GlcNAc transporter that delivers an essential substrate for the maturation of N-glycans and the GIPC class of sphingolipids.},
doi = {10.1038/s41477-018-0235-5},
journal = {Nature Plants},
issn = {2055-0278},
number = 10,
volume = 4,
place = {United States},
year = {2018},
month = {9}
}

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