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Biosynthesis of UDP-xylose: characterization of membrane-bound AtUxs2

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Abstract

UDP-xylose (UDP-Xyl) is a sugar donor for the synthesis of glycoproteins, polysaccharides, various metabolites, and oligosaccharides in plants, vertebrates, and fungi. In plants, the biosynthesis of UDP-Xyl from UDP-glucuronic acid (UDP-GlcA) appears to be catalyzed by numerous UDP-glucuronic acid decarboxylase (Uxs) isoforms. For example, six Uxs isoforms in Arabidopsis thaliana (L.) and four in rice have been identified. However, the reason/s for the existence of several isoforms that are necessary for the synthesis of UDP-Xyl remains unknown. Here, we describe a Uxs isoform in Arabidopsis, AtUXS2, encoding an integral membrane protein that appears to be localized to the Golgi apparatus. The enzyme is a dimer and has distinct properties. Unlike the UXS3 isoform, which is shown here to be a soluble protein, the UXS2 isoform is membrane bound. The characteristics of the membrane-bound AtUxs2 and cytosolic AtUxs3 support the hypothesis that unique UDP-GlcA-DCs possessing distinct sub-cellular localizations can spatially regulate specific xylosylation events in plant cells.

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Abbreviations

UDP-Glc:

UDP-glucose

UDP-Xyl:

UDP-xylose

UDP-GlcA:

UDP-glucuronic acid

DTT:

Dithiothrietol

UDP-GlcA-DC:

UDP-glucuronic acid decarboxylase

PMSF:

Phenylmethylsulfonyl fluoride

PCR:

Polymerase chain reaction

RT:

Reverse transcription

IPTG:

Isopropyl-δ-D-thiogalactoside-EST expressed sequence tag

UXS:

UDP-Xyl Synthase

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Acknowledgements

We wish to thank Dr. Aki Nakano for the AtSed5 and AtRer1B constructs and Kristen Herrick and Praveen Polavarapu for their assistance in cloning. We also wish to thank Dr. John Glushka of the CCRC for his expertise and extensive discussion regarding various aspects of NMR data, and Dr. Michael Hahn for his constructive comments on this manuscript. This research was supported in part by U.S. Department of Agriculture grant 2002-35318-12620 (to MB-P) and by a U.S. Department of Energy-funded Center grant (DE-FG05-93ER20097) for Plant and Microbial Complex Carbohydrates.

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  1. Complex Carbohydrate Research Center and Department of Plant Biology, University of Georgia, 315 Riverbend Road, Athens, GA, 30602-4712, USA

    Sivakumar Pattathil, April D. Harper & Maor Bar-Peled

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  1. Sivakumar Pattathil
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  2. April D. Harper
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  3. Maor Bar-Peled
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Correspondence to Maor Bar-Peled.

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Pattathil, S., Harper, A.D. & Bar-Peled, M. Biosynthesis of UDP-xylose: characterization of membrane-bound AtUxs2. Planta 221, 538–548 (2005). https://doi.org/10.1007/s00425-004-1471-7

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  • DOI: https://doi.org/10.1007/s00425-004-1471-7

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