The human solute carrier gene SLC35B4 encodes a bifunctional nucleotide sugar transporter with specificity for UDP-xylose and UDP-N-acetylglucosamine.
The transport of nucleotide sugars from the cytoplasm into the Golgi apparatus is mediated by specialized type III proteins, the nucleotide sugar transporters (NSTs). Transport assays carried out in vitro with Golgi vesicles from mammalian cells showed specific uptake for a total of eight nucleotide sugars. When this study was started, NSTs with transport activities for all but two nucleotide sugars (UDP-Xyl and UDP-Glc) had been cloned. Aiming at identifying these elusive NSTs, bioinformatic methods were used to display putative NST sequences in the human genome. Ten open reading frames were identified, cloned, and heterologously expressed in yeast. Transport capabilities for UDP-Glc and UDP-Xyl were determined with Golgi vesicles isolated from transformed cells. Although a potential UDP-Glc transporter could not be identified due to the high endogenous transport background, the measurement of UDP-Xyl transport was possible on a zero background. Vesicles from yeast cells expressing the human gene SLC35B4 showed specific uptake of UDP-Xyl, and subsequent testing of other nucleotide sugars revealed a second activity for UDP-GlcNAc. Expression of the epitope-tagged SLC35B4 in mammalian cells demonstrated strict Golgi localization. Because decarboxylation of UDP-GlcA is known to produce UDP-Xyl directly in the endoplasmic reticulum and Golgi lumen, our data demonstrate that two ways exist to deliver UDP-Xyl to the Golgi apparatus.
Ashikov
Angel
A
Zelluläre Chemie, Zentrum Biochemie, Medizinische Hochschule Hannover, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
Routier
Françoise
F
Fuhlrott
Jutta
J
Helmus
Yvonne
Y
Wild
Martin
M
Gerardy-Schahn
Rita
R
Bakker
Hans
H
D001483:Base Sequence; D001692:Biological Transport; D002460:Cell Line; D019295:Computational Biology; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000502:physiology; D033704:Nucleotide Transport Proteins / Q000235:genetics* / Q000502:physiology; D012441:Saccharomyces cerevisiae; D013379:Substrate Specificity; D014162:Transfection; D014170:Transformation, Genetic; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism*; D014540:Uridine Diphosphate Xylose / Q000378:metabolism*
D009004:Monosaccharide Transport Proteins; D033704:Nucleotide Transport Proteins; C503492:SLC35B4 protein, human; D014540:Uridine Diphosphate Xylose; D014537:Uridine Diphosphate N-Acetylglucosamine