Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1996 Dec;112(4):1585–1594. doi: 10.1104/pp.112.4.1585

Evidence for a UDP-Glucose Transporter in Golgi Apparatus-Derived Vesicles from Pea and Its Possible Role in Polysaccharide Biosynthesis.

P Munoz 1, L Norambuena 1, A Orellana 1
PMCID: PMC158091  PMID: 12226465

Abstract

The Golgi apparatus in plant cells is involved in hemicellulose and pectin biosynthesis. While it is known that glucan synthase I is responsible for the formation of [beta]-l-4-linked glucose (Glc) polymers and uses UDP-Glc as a substrate, very little is known about the topography of reactions leading to the biosynthesis of polysaccharides in this organelle. We isolated from pea (Pisum sativum) stems a fraction highly enriched in Golgi apparatus-derived vesicles that are sealed and have the same topographical orientation that the membranes have in vivo. Using these vesicles and UDP-Glc, we reconstituted polysaccharide biosynthesis in vitro and found evidence for a luminal location of the active site of glucan synthase I. In addition, we identified a UDP-Glc transport activity, which is likely to be involved in supplying substrate for glucan synthase I. We found that UDP-Glc transport is protein mediated. Moreover, our results suggest that UDP-Glc transport is coupled to the exit of a luminal uridine-containing nucleotide via an antiporter mechanism. We suggest that UDP-Glc is transported into the lumen of Golgi and that Glc is transferred to a polysaccharide chain, whereas the nucleotide moiety leaves the vesicle by an antiporter mechanism.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abdul-Baki A. A., Ray P. M. Regulation by auxin of carbohydrate metabolism involved in cell wall synthesis by pea stem tissue. Plant Physiol. 1971 Apr;47(4):537–544. doi: 10.1104/pp.47.4.537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abeijon C., Orlean P., Robbins P. W., Hirschberg C. B. Topography of glycosylation in yeast: characterization of GDPmannose transport and lumenal guanosine diphosphatase activities in Golgi-like vesicles. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6935–6939. doi: 10.1073/pnas.86.18.6935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Abeijon C., Yanagisawa K., Mandon E. C., Häusler A., Moremen K., Hirschberg C. B., Robbins P. W. Guanosine diphosphatase is required for protein and sphingolipid glycosylation in the Golgi lumen of Saccharomyces cerevisiae. J Cell Biol. 1993 Jul;122(2):307–323. doi: 10.1083/jcb.122.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berninsone P., Miret J. J., Hirschberg C. B. The Golgi guanosine diphosphatase is required for transport of GDP-mannose into the lumen of Saccharomyces cerevisiae Golgi vesicles. J Biol Chem. 1994 Jan 7;269(1):207–211. [PubMed] [Google Scholar]
  5. Brandan E., Fleischer B. Orientation and role of nucleosidediphosphatase and 5'-nucleotidase in Golgi vesicles from rat liver. Biochemistry. 1982 Sep 14;21(19):4640–4645. doi: 10.1021/bi00262a019. [DOI] [PubMed] [Google Scholar]
  6. Capasso J. M., Hirschberg C. B. 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. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7051–7055. doi: 10.1073/pnas.81.22.7051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dauwalder M., Whaley W. G., Kephart J. E. Phosphatases and differentiation of the Golgi apparatus. J Cell Sci. 1969 Mar;4(2):455–497. doi: 10.1242/jcs.4.2.455. [DOI] [PubMed] [Google Scholar]
  8. Dhugga K. S., Ray P. M. Purification of 1,3-beta-D-glucan synthase activity from pea tissue. Two polypeptides of 55 kDa and 70 kDa copurify with enzyme activity. Eur J Biochem. 1994 Mar 15;220(3):943–953. doi: 10.1111/j.1432-1033.1994.tb18698.x. [DOI] [PubMed] [Google Scholar]
  9. Dhugga K. S., Ulvskov P., Gallagher S. R., Ray P. M. Plant polypeptides reversibly glycosylated by UDP-glucose. Possible components of Golgi beta-glucan synthase in pea cells. J Biol Chem. 1991 Nov 15;266(32):21977–21984. [PubMed] [Google Scholar]
  10. Driouich A., Faye L., Staehelin L. A. The plant Golgi apparatus: a factory for complex polysaccharides and glycoproteins. Trends Biochem Sci. 1993 Jun;18(6):210–214. doi: 10.1016/0968-0004(93)90191-o. [DOI] [PubMed] [Google Scholar]
  11. Gibeaut D. M., Carpita N. C. Biosynthesis of plant cell wall polysaccharides. FASEB J. 1994 Sep;8(12):904–915. doi: 10.1096/fasebj.8.12.8088456. [DOI] [PubMed] [Google Scholar]
  12. Hayashi T., Koyama T., Matsuda K. Formation of UDP-Xylose and Xyloglucan in Soybean Golgi Membranes. Plant Physiol. 1988 Jun;87(2):341–345. doi: 10.1104/pp.87.2.341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hayashi T., Maclachlan G. Biosynthesis of pentosyl lipids by pea membranes. Biochem J. 1984 Feb 1;217(3):791–803. doi: 10.1042/bj2170791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hayashi T., Matsuda K. Biosynthesis of xyloglucan in suspension-cultured soybean cells. Occurrence and some properties of xyloglucan 4-beta-D-glucosyltransferase and 6-alpha-D-xylosyltransferase. J Biol Chem. 1981 Nov 10;256(21):11117–11122. [PubMed] [Google Scholar]
  15. Hirschberg C. B., Snider M. D. Topography of glycosylation in the rough endoplasmic reticulum and Golgi apparatus. Annu Rev Biochem. 1987;56:63–87. doi: 10.1146/annurev.bi.56.070187.000431. [DOI] [PubMed] [Google Scholar]
  16. Mitsui T., Honma M., Kondo T., Hashimoto N., Kimura S., Igaue I. Structure and Function of the Golgi Complex in Rice Cells (II. Purification and Characterization of Golgi Membrane-Bound Nucleoside Diphosphatase). Plant Physiol. 1994 Sep;106(1):119–125. doi: 10.1104/pp.106.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. PALADINI A. C., LELOIR L. F. Studies on uridine-diphosphate-glucose. Biochem J. 1952 Jun;51(3):426–430. doi: 10.1042/bj0510426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ray P. M., Shininger T. L., Ray M. M. ISOLATION OF beta-GLUCAN SYNTHETASE PARTICLES FROM PLANT CELLS AND IDENTIFICATION WITH GOLGI MEMBRANES. Proc Natl Acad Sci U S A. 1969 Oct;64(2):605–612. doi: 10.1073/pnas.64.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Waldman B. C., Rudnick G. UDP-GlcNAc transport across the Golgi membrane: electroneutral exchange for dianionic UMP. Biochemistry. 1990 Jan 9;29(1):44–52. doi: 10.1021/bi00453a006. [DOI] [PubMed] [Google Scholar]
  20. White A. R., Xin Y., Pezeshk V. Xyloglucan glucosyltransferase in Golgi membranes from Pisum sativum (pea). Biochem J. 1993 Aug 15;294(Pt 1):231–238. doi: 10.1042/bj2940231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zhang G. F., Staehelin L. A. Functional compartmentation of the Golgi apparatus of plant cells : immunocytochemical analysis of high-pressure frozen- and freeze-substituted sycamore maple suspension culture cells. Plant Physiol. 1992 Jul;99(3):1070–1083. doi: 10.1104/pp.99.3.1070. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES