Transport of UDP-galactose in plants -: Identification and functional characterization of AtUTr1, an Arabidopsis thaliana UDP-galactose/UDP-glucose transporter

被引:74
作者
Norambuena, L
Marchant, L
Berninsone, P
Hirschberg, CB
Silva, H
Orellana, A
机构
[1] Univ Chile, Dept Biol, Fac Sci, Santiago, Chile
[2] Univ Chile, Millenium Inst Cell Biol & Biotechnol, Santiago, Chile
[3] Boston Univ, Sch Dent Med, Dept Mol & Cell Biol, Boston, MA 02118 USA
关键词
D O I
10.1074/jbc.M204081200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The synthesis of non-cellulosic polysaccharides and glycoproteins in the plant cell Golgi apparatus requires UDP-galactose as substrate. The topology of these reactions is not known, although the orientation of a plant galactosyltransferase involved in the biosynthesis of galactomannans in fenugreek is consistent with a requirement for UDP-galactose in the lumen of the Golgi cisternae. Here we provide evidence that sealed, right-sideout Golgi vesicles isolated from pea stems transport UDP-galactose into their lumen and transfer galactose, likely to polysaccharides and other acceptors. In addition, we identified and cloned AtUTr1, a gene from Arabidopsis thaliana that encodes a multitransmembrane hydrophobic protein similar to nucleotide sugar transporters. Northern analysis showed that AtUTr1 is indeed expressed in Arabidopsis. AtUTr1 is able to complement the phenotype of MDCK ricin-resistant cells; a mammalian cell line deficient in transport of UDP-galactose into the Golgi. In vitro assays using a Golgi-enriched vesicle fraction obtained from Saccharomyces cerevisiae expressing AtUTr1-MycHis is able to transport UDP-galactose but also UDP-glucose. AtUTr1-MycHis does not transport GDP-mannose, GDP-fucose, CMP-sialic acid, UDP-glucuronic acid, or UDP-xylose when expressed in S. cerevisiae. AtUTr1 is the first transporter described that is able to transport UDP-galactose and UDP-glucose. Thus AtUTr1 may play an important role in the synthesis of glycoconjugates in Arabidopsis that contain galactose and glucose.
引用
收藏
页码:32923 / 32929
页数:7
相关论文
共 38 条
[1]   TOPOGRAPHY OF GLYCOSYLATION IN YEAST - CHARACTERIZATION OF GDPMANNOSE TRANSPORT AND LUMENAL GUANOSINE DIPHOSPHATASE ACTIVITIES IN GOLGI-LIKE VESICLES [J].
ABEIJON, C ;
ORLEAN, P ;
ROBBINS, PW ;
HIRSCHBERG, CB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (18) :6935-6939
[2]  
Baldwin TC, 2001, PLANT CELL, V13, P2283, DOI 10.1105/tpc.13.10.2283
[3]   SQV-7, a protein involved in Caenorhabditis elegans epithelial invagination and early embryogenesis, transports UDP-glucuronic acid, UDP-N-acetylgalactosamine, and UDP-galactose [J].
Berninsone, P ;
Hwang, HY ;
Zemtseva, I ;
Horvitz, HR ;
Hirschberg, CB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (07) :3738-3743
[4]   Functional expression of the murine gels CMP-sialic acid transporter in Saccharomyces cerevisiae [J].
Berninsone, P ;
Eckhardt, M ;
GerardySchahn, R ;
Hirschberg, CB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (19) :12616-12619
[5]   Nucleotide sugar transporters of the Golgi apparatus [J].
Berninsone, PM ;
Hirschberg, CB .
CURRENT OPINION IN STRUCTURAL BIOLOGY, 2000, 10 (05) :542-547
[6]  
BRANDLI AW, 1988, J BIOL CHEM, V263, P16283
[7]   MECHANISMS OF GLYCOSYLATION AND SULFATION IN THE GOLGI-APPARATUS - EVIDENCE FOR NUCLEOTIDE SUGAR NUCLEOSIDE MONOPHOSPHATE AND NUCLEOTIDE SULFATE NUCLEOSIDE MONOPHOSPHATE ANTIPORTS IN THE GOLGI-APPARATUS MEMBRANE [J].
CAPASSO, JM ;
HIRSCHBERG, CB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (22) :7051-7055
[8]  
CAPASSO JM, 1984, J BIOL CHEM, V259, P4263
[9]  
Carpita N., 2000, BIOCH MOL BIOL PLANT, P52
[10]   A SPECIALIZED PATHWAY AFFECTING VIRULENCE GLYCOCONJUGATES OF LEISHMANIA [J].
DESCOTEAUX, A ;
LUO, Y ;
TURCO, SJ ;
BEVERLEY, SM .
SCIENCE, 1995, 269 (5232) :1869-1872