Glycobiology Advance Access originally published online on September 29, 2004
Glycobiology 2005 15(2):193-201; doi:10.1093/glycob/cwh159
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Glycobiology vol. 15 no. 2 © Oxford University Press 2005; all rights reserved.
Molecular cloning of two Arabidopsis UDP-galactose transporters by complementation of a deficient Chinese hamster ovary cell line
3 Plant Research International, Wageningen University and Research Centre, P.O.Box 16, 6700AA, Wageningen, The Netherlands; 4 Zelluläre Chemie, Medizinische Hochschule Hannover, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany; and 5 Institute of Food Safety (RIKILT), Wageningen University and Research Centre, P.O. Box 230, 6700AE Wageningen, The Netherlands
2 To whom correspondence should be addressed; e-mail: bakker.hans{at}mh-hannover.de
Received on July 16, 2004; revised on September 21, 2004; accepted on September 23, 2004
Nucleotide-sugar transporters (NSTs) form a family of structurally related transmembrane proteins that transport nucleotide-sugars from the cytoplasm to the endoplasmic reticulum and Golgi lumen. In these organelles, activated sugars are substrates for various glycosyltransferases involved in oligo- and polysaccharide biosynthesis. The Arabidopsis thaliana genome contains more than 40 members of this transporter gene family, of which only a few are functionally characterized. In this study, two Arabidopsis UDP-galactose transporter cDNAs (UDP-GalT1 and UDP-GalT2) are isolated by expression cloning using a Chinese hamster ovary cell line (CHO-Lec8) deficient in UDP-galactose transport. The isolated genes show only 21% identity to each other and very limited sequence identity with human and yeast UDP-galactose transporters and other NSTs. Despite this low overall identity, the two proteins clearly belong to the same gene family. Besides complementing Lec8 cells, the two NSTs are shown to transport exclusively UDP-galactose by an in vitro NST assay. The most homologous proteins with known function are plant transporters that locate in the inner chloroplast membrane and transport triose-phosphate, phosphoenolpyruvate, glucose-6-phosphate, and xylulose 5-phosphate. Also, the latter proteins are members of the same family, which therefore has been named the NST/triose-phosphate transporter family.
Key words: cell wall / expression cloning / Golgi / membrane / nucleotide-sugar transporter
1 These authors contributed equally to this work.
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