skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development

Abstract

Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rhamnogalacturonan II. Furthermore, these lines display reduced L-fucose content in N-glycan structures accompanied by severe developmental growth defects. We conclude that GFT1 is the major nucleotide sugar transporter for import of GDP-L-fucose into the Golgi and is required for proper plant growth and development.

Authors:
 [1];  [2];  [3];  [4];  [4];  [3];  [5];  [6];  [1]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Joint BioEnergy Inst.; Univ. of Melbourne (Australia). ARC Centre of Excellence in Plant Cell Walls
  2. Univ. of Melbourne (Australia). ARC Centre of Excellence in Plant Cell Walls; Univ. of Copenhagen (Denmark). Dept. of Plant and Environmental Sciences
  3. Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology
  4. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Joint BioEnergy Inst.
  5. Andres Bello National Univ., Santiago (Chile). Biotechnology Center; Center for Genome Regulation, Santiago (Chile)
  6. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Joint BioEnergy Inst.; Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology
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:
1379504
Grant/Contract Number:
AC02-05CH11231
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 7; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; plant development; plant transporters

Citation Formats

Rautengarten, Carsten, Ebert, Berit, Liu, Lifeng, Stonebloom, Solomon, Smith-Moritz, Andreia M., Pauly, Markus, Orellana, Ariel, Scheller, Henrik Vibe, and Heazlewood, Joshua L. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development. United States: N. p., 2016. Web. doi:10.1038/ncomms12119.
Rautengarten, Carsten, Ebert, Berit, Liu, Lifeng, Stonebloom, Solomon, Smith-Moritz, Andreia M., Pauly, Markus, Orellana, Ariel, Scheller, Henrik Vibe, & Heazlewood, Joshua L. The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development. United States. doi:10.1038/ncomms12119.
Rautengarten, Carsten, Ebert, Berit, Liu, Lifeng, Stonebloom, Solomon, Smith-Moritz, Andreia M., Pauly, Markus, Orellana, Ariel, Scheller, Henrik Vibe, and Heazlewood, Joshua L. 2016. "The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development". United States. doi:10.1038/ncomms12119. https://www.osti.gov/servlets/purl/1379504.
@article{osti_1379504,
title = {The Arabidopsis Golgi-localized GDP-L-fucose transporter is required for plant development},
author = {Rautengarten, Carsten and Ebert, Berit and Liu, Lifeng and Stonebloom, Solomon and Smith-Moritz, Andreia M. and Pauly, Markus and Orellana, Ariel and Scheller, Henrik Vibe and Heazlewood, Joshua L.},
abstractNote = {Nucleotide sugar transport across Golgi membranes is essential for the luminal biosynthesis of glycan structures. Here we identify GDP-fucose transporter 1 (GFT1), an Arabidopsis nucleotide sugar transporter that translocates GDP-L-fucose into the Golgi lumen. Using proteo-liposome-based transport assays, we show that GFT preferentially transports GDP-L-fucose over other nucleotide sugars in vitro, while GFT1-silenced plants are almost devoid of L-fucose in cell wall-derived xyloglucan and rhamnogalacturonan II. Furthermore, these lines display reduced L-fucose content in N-glycan structures accompanied by severe developmental growth defects. We conclude that GFT1 is the major nucleotide sugar transporter for import of GDP-L-fucose into the Golgi and is required for proper plant growth and development.},
doi = {10.1038/ncomms12119},
journal = {Nature Communications},
number = ,
volume = 7,
place = {United States},
year = 2016,
month = 7
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Save / Share:
  • UDP-glucuronic acid (UDP-GlcA) is the precursor of many plant cell wall polysaccharides and is required for production of seed mucilage. Following synthesis in the cytosol, it is transported into the lumen of the Golgi apparatus, where it is converted to UDP-galacturonic acid (UDP-GalA), UDP-arabinose, and UDP-xylose. To identify the Golgi-localized UDP-GlcA transporter, we screened Arabidopsis thaliana mutants in genes coding for putative nucleotide sugar transporters for altered seed mucilage, a structure rich in the GalA-containing polysaccharide rhamnogalacturonan I. As a result, we identified UUAT1, which encodes a Golgi-localized protein that transports UDP-GlcA and UDP-GalA in vitro. The seed coat ofmore » uuat1 mutants had less GalA, rhamnose, and xylose in the soluble mucilage, and the distal cell walls had decreased arabinan content. Cell walls of other organs and cells had lower arabinose levels in roots and pollen tubes, but no differences were observed in GalA or xylose contents. Furthermore, the GlcA content of glucuronoxylan in the stem was not affected in the mutant. Interestingly, the degree of homogalacturonan methylation increased in uuat1. These results suggest that this UDP-GlcA transporter plays a key role defining the seed mucilage sugar composition and that its absence produces pleiotropic effects in this component of the plant extracellular matrix.« less
  • Male Sterile2 (MS2) is predicted to encode a fatty acid reductase required for pollen wall development in Arabidopsis (Arabidopsis thaliana). Transient expression of MS2 in tobacco (Nicotiana benthamiana) leaves resulted in the accumulation of significant levels of C16 and C18 fatty alcohols. Expression of MS2 fused with green fluorescent protein revealed that an amino-terminal transit peptide targets the MS2 to plastids. The plastidial localization of MS2 is biologically important because genetic complementation of MS2 in ms2 homozygous plants was dependent on the presence of its amino-terminal transit peptide or that of the Rubisco small subunit protein amino-terminal transit peptide. Inmore » addition, two domains, NAD(P)H-binding domain and sterile domain, conserved in MS2 and its homologs were also shown to be essential for MS2 function in pollen exine development by genetic complementation testing. Direct biochemical analysis revealed that purified recombinant MS2 enzyme is able to convert palmitoyl-Acyl Carrier Protein to the corresponding C16:0 alcohol with NAD(P)H as the preferred electron donor. Using optimized reaction conditions (i.e. at pH 6.0 and 30 C), MS2 exhibits a K{sub m} for 16:0-Acyl Carrier Protein of 23.3 {+-} 4.0 {mu}m, a V{sub max} of 38.3 {+-} 4.5 nmol mg{sup -1} min{sup -1}, and a catalytic efficiency/K{sub m} of 1,873 m{sup -1} s{sup -1}. Based on the high homology of MS2 to other characterized fatty acid reductases, it was surprising that MS2 showed no activity against palmitoyl- or other acyl-coenzyme A; however, this is consistent with its plastidial localization. In summary, genetic and biochemical evidence demonstrate an MS2-mediated conserved plastidial pathway for the production of fatty alcohols that are essential for pollen wall biosynthesis in Arabidopsis.« less
  • 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 majoritymore » 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.« less