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 Doñas-Cofré, Adrián Moreno, Susana Saéz-Aguayo, Francisca Blanco, Jennifer C. Mortimer, Alex Schultink, Wolf Dieter Reiter, Paul Dupree, Markus Pauly, Joshua L. Heazlewood, Henrik V. Scheller, Ariel Orellana

Resultado de la investigación: Article

46 Citas (Scopus)

Resumen

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.

Idioma originalEnglish
Páginas (desde-hasta)11563-11568
Número de páginas6
PublicaciónProceedings of the National Academy of Sciences of the United States of America
Volumen111
N.º31
DOI
EstadoPublished - 5 ago 2014

Huella dactilar

Rhamnose
Uridine Diphosphate
Arabidopsis
Nucleotides
Cell Wall
Polysaccharides
Liposomes
Cytosol
Uridine Diphosphate Galactose
Plant Cells
Golgi Apparatus
UDP-galactose translocator
Galactose
Mass Spectrometry
Growth

ASJC Scopus subject areas

  • General

Citar esto

Rautengarten, Carsten ; Ebert, Berit ; Moreno, Ignacio ; Temple, Henry ; Herter, Thomas ; Link, Bruce ; Doñas-Cofré, Daniela ; Moreno, Adrián ; Saéz-Aguayo, Susana ; Blanco, Francisca ; Mortimer, Jennifer C. ; Schultink, Alex ; Reiter, Wolf Dieter ; Dupree, Paul ; Pauly, Markus ; Heazlewood, Joshua L. ; Scheller, Henrik V. ; Orellana, Ariel. / The Golgi localized bifunctional UDP-rhamnose/UDP-galactose transporter family of Arabidopsis. En: Proceedings of the National Academy of Sciences of the United States of America. 2014 ; Vol. 111, N.º 31. pp. 11563-11568.
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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.",
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Rautengarten, C, Ebert, B, Moreno, I, Temple, H, Herter, T, Link, B, Doñas-Cofré, D, Moreno, A, Saéz-Aguayo, S, Blanco, F, Mortimer, JC, Schultink, A, Reiter, WD, 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, vol. 111, n.º 31, pp. 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; Temple, Henry; Herter, Thomas; Link, Bruce; Doñas-Cofré, Daniela; Moreno, Adrián; Saéz-Aguayo, Susana; Blanco, Francisca; Mortimer, Jennifer C.; Schultink, Alex; Reiter, Wolf Dieter; Dupree, Paul; Pauly, Markus; Heazlewood, Joshua L.; Scheller, Henrik V.; Orellana, Ariel.

En: Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, N.º 31, 05.08.2014, p. 11563-11568.

Resultado de la investigación: Article

TY - JOUR

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 - Doñas-Cofré, Daniela

AU - Moreno, Adrián

AU - Saéz-Aguayo, 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.

KW - Galactan

KW - Glycan biosynthesis

KW - Membrane transport

KW - Proteoliposomes

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U2 - 10.1073/pnas.1406073111

DO - 10.1073/pnas.1406073111

M3 - Article

C2 - 25053812

AN - SCOPUS:84905661578

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

SN - 0027-8424

IS - 31

ER -