The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis

Carsten Rautengarten; Berit Ebert; Ignacio Moreno; Henry Temple; Thomas Herter; Bruce Link; Daniela Donas-Cofre; Adrian Moreno; Susana Saez-Aguayoc; Francisca Blanco; Jennifer C. Mortimer; Alex Schultink; Wolf-Dieter Reiter; Paul Dupree; Markus Pauly; Joshua L. Heazlewood; Henrik V. Scheller; Ariel Orellana

doi:10.1073/pnas.1406073111

The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis

Carsten Rautengarten, Berit Ebert, Ignacio Moreno, Henry Temple, Thomas Herter, Bruce Link, Daniela Donas-Cofre, Adrian Moreno, Susana Saez-Aguayoc, Francisca Blanco, Jennifer C. Mortimer, Alex Schultink, Wolf-Dieter Reiter, Paul Dupree, Markus Pauly, Joshua L. Heazlewood, Henrik V. Scheller, Ariel Orellana

Planteglycobiologi

64 Citationer (Scopus)

Abstract

Delivery of nucleotide sugar substrates into the Golgi apparatus and endoplasmic reticulum for processes such as cell wall biosynthesis and protein glycosylation is critical for plant growth and development. Plant genomes encode large families of uncharacterized nucleotide sugar transporters that are specifically presumed to deliver the diverse array of nucleotide sugars found in plants. This study has developed a novel approach that enabled functional characterization of six bifunctional UDP-rhamnose (Rha)/UDP-galactose (Gal) transporters from Arabidopsis. An analysis of loss-of-function and overexpression lines for two of these transporters identified biochemical alterations supporting their roles in the biosynthesis of Rha- and Gal-containing polysaccharides. Thus, cell wall polysaccharide biosynthesis in the Golgi apparatus of plants is likely also regulated by substrate transport mechanisms.

Originalsprog	Engelsk
Tidsskrift	Proceedings of the National Academy of Sciences of the United States of America
Vol/bind	111
Udgave nummer	31
Sider (fra-til)	11563-11568
Antal sider	6
ISSN	0027-8424
DOI	https://doi.org/10.1073/pnas.1406073111
Status	Udgivet - 5 aug. 2014

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10.1073/pnas.1406073111

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Rautengarten, C., Ebert, B., Moreno, I., Temple, H., Herter, T., Link, B., Donas-Cofre, D., Moreno, A., Saez-Aguayoc, S., Blanco, F., Mortimer, J. C., Schultink, A., Reiter, W-D., Dupree, P., Pauly, M., Heazlewood, J. L., Scheller, H. V., & Orellana, A. (2014). The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America, 111(31), 11563-11568. https://doi.org/10.1073/pnas.1406073111

The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis. / Rautengarten, Carsten; Ebert, Berit; Moreno, Ignacio et al.

I: Proceedings of the National Academy of Sciences of the United States of America, Bind 111, Nr. 31, 05.08.2014, s. 11563-11568.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

Rautengarten, C, Ebert, B, Moreno, I, Temple, H, Herter, T, Link, B, Donas-Cofre, D, Moreno, A, Saez-Aguayoc, S, Blanco, F, Mortimer, JC, Schultink, A, Reiter, W-D, Dupree, P, Pauly, M, Heazlewood, JL, Scheller, HV & Orellana, A 2014, 'The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis', Proceedings of the National Academy of Sciences of the United States of America, bind 111, nr. 31, s. 11563-11568. https://doi.org/10.1073/pnas.1406073111

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title = "The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis",

abstract = "Plant cells are surrounded by a cell wall that plays a key role in plant growth, structural integrity, and defense. The cell wall is a complex and diverse structure that is mainly composed of polysaccharides. The majority of noncellulosic cell wall polysaccharides are produced in the Golgi apparatus from nucleotide sugars that are predominantly synthesized in the cytosol. The transport of these nucleotide sugars from the cytosol into the Golgi lumen is a critical process for cell wall biosynthesis and is mediated by a family of nucleotide sugar transporters (NSTs). Numerous studies have sought to characterize substrate-specific transport by NSTs; however, the availability of certain substrates and a lack of robust methods have proven problematic. Consequently, we have developed a novel approach that combines reconstitution of NSTs into liposomes and the subsequent assessment of nucleotide sugar uptake by mass spectrometry. To address the limitation of substrate availability, we also developed a two-step reaction for the enzymatic synthesis of UDP-L-rhamnose (Rha) by expressing the two active domains of the Arabidopsis UDP-L-Rha synthase. The liposome approach and the newly synthesized substrates were used to analyze a clade of Arabidopsis NSTs, resulting in the identification and characterization of six bifunctional UDP-L-Rha/UDP-D-galactose (Gal) transporters (URGTs). Further analysis of loss-of-function and overexpression plants for two of these URGTs supported their roles in the transport of UDP-L-Rha and UDP-D-Gal for matrix polysaccharide biosynthesis.",

author = "Carsten Rautengarten and Berit Ebert and Ignacio Moreno and Henry Temple and Thomas Herter and Bruce Link and Daniela Donas-Cofre and Adrian Moreno and Susana Saez-Aguayoc and Francisca Blanco and Mortimer, {Jennifer C.} and Alex Schultink and Wolf-Dieter Reiter and Paul Dupree and Markus Pauly and Heazlewood, {Joshua L.} and Scheller, {Henrik V.} and Ariel Orellana",

year = "2014",

month = aug,

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doi = "10.1073/pnas.1406073111",

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T1 - The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis

AU - Rautengarten, Carsten

AU - Ebert, Berit

AU - Moreno, Ignacio

AU - Temple, Henry

AU - Herter, Thomas

AU - Link, Bruce

AU - Donas-Cofre, Daniela

AU - Moreno, Adrian

AU - Saez-Aguayoc, Susana

AU - Blanco, Francisca

AU - Mortimer, Jennifer C.

AU - Schultink, Alex

AU - Reiter, Wolf-Dieter

AU - Dupree, Paul

AU - Pauly, Markus

AU - Heazlewood, Joshua L.

AU - Scheller, Henrik V.

AU - Orellana, Ariel

PY - 2014/8/5

Y1 - 2014/8/5

N2 - Plant cells are surrounded by a cell wall that plays a key role in plant growth, structural integrity, and defense. The cell wall is a complex and diverse structure that is mainly composed of polysaccharides. The majority of noncellulosic cell wall polysaccharides are produced in the Golgi apparatus from nucleotide sugars that are predominantly synthesized in the cytosol. The transport of these nucleotide sugars from the cytosol into the Golgi lumen is a critical process for cell wall biosynthesis and is mediated by a family of nucleotide sugar transporters (NSTs). Numerous studies have sought to characterize substrate-specific transport by NSTs; however, the availability of certain substrates and a lack of robust methods have proven problematic. Consequently, we have developed a novel approach that combines reconstitution of NSTs into liposomes and the subsequent assessment of nucleotide sugar uptake by mass spectrometry. To address the limitation of substrate availability, we also developed a two-step reaction for the enzymatic synthesis of UDP-L-rhamnose (Rha) by expressing the two active domains of the Arabidopsis UDP-L-Rha synthase. The liposome approach and the newly synthesized substrates were used to analyze a clade of Arabidopsis NSTs, resulting in the identification and characterization of six bifunctional UDP-L-Rha/UDP-D-galactose (Gal) transporters (URGTs). Further analysis of loss-of-function and overexpression plants for two of these URGTs supported their roles in the transport of UDP-L-Rha and UDP-D-Gal for matrix polysaccharide biosynthesis.

AB - Plant cells are surrounded by a cell wall that plays a key role in plant growth, structural integrity, and defense. The cell wall is a complex and diverse structure that is mainly composed of polysaccharides. The majority of noncellulosic cell wall polysaccharides are produced in the Golgi apparatus from nucleotide sugars that are predominantly synthesized in the cytosol. The transport of these nucleotide sugars from the cytosol into the Golgi lumen is a critical process for cell wall biosynthesis and is mediated by a family of nucleotide sugar transporters (NSTs). Numerous studies have sought to characterize substrate-specific transport by NSTs; however, the availability of certain substrates and a lack of robust methods have proven problematic. Consequently, we have developed a novel approach that combines reconstitution of NSTs into liposomes and the subsequent assessment of nucleotide sugar uptake by mass spectrometry. To address the limitation of substrate availability, we also developed a two-step reaction for the enzymatic synthesis of UDP-L-rhamnose (Rha) by expressing the two active domains of the Arabidopsis UDP-L-Rha synthase. The liposome approach and the newly synthesized substrates were used to analyze a clade of Arabidopsis NSTs, resulting in the identification and characterization of six bifunctional UDP-L-Rha/UDP-D-galactose (Gal) transporters (URGTs). Further analysis of loss-of-function and overexpression plants for two of these URGTs supported their roles in the transport of UDP-L-Rha and UDP-D-Gal for matrix polysaccharide biosynthesis.

U2 - 10.1073/pnas.1406073111

DO - 10.1073/pnas.1406073111

M3 - Journal article

C2 - 25053812

SN - 0027-8424

VL - 111

SP - 11563

EP - 11568

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 31

ER -

The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis

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