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The gene defective in leukocyte adhesion deficiency II encodes a putative GDP-fucose transporter

Nature Genetics volume 28, pages 69–72 (2001)Cite this article

Abstract

Leukocyte adhesion deficiency II (LAD II) is characterized by the lack of fucosylated glycoconjugates, including selectin ligands, causing immunodeficiency and severe mental and growth retardation1,2,3. No deficiency in fucosyltransferase activities2,4 or in the activities of enzymes involved in GDP-fucose biosynthesis5 has been found. Instead, the transport of GDP-fucose into isolated Golgi vesicles of LAD II cells appeared to be reduced6. To identify the gene mutated in LAD II, we cloned 12 cDNAs from Caenorhabditis elegans, encoding multi-spanning transmembrane proteins with homology to known nucleotide sugar transporters, and transfected them into fibroblasts from an LAD II patient. One of these clones re-established expression of fucosylated glycoconjugates with high efficiency and allowed us to identify a human homolog with 55% identity, which also directed re-expression of fucosylated glycoconjugates. Both proteins were localized to the Golgi. The corresponding endogenous protein in LAD II cells had an R147C amino acid change in the conserved fourth transmembrane region. Overexpression of this mutant protein in cells from a patient with LAD II did not rescue fucosylation, demonstrating that the point mutation affected the activity of the protein. Thus, we have identified the first putative GDP-fucose transporter, which has been highly conserved throughout evolution. A point mutation in its gene is responsible for the disease in this patient with LAD II.

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Figure 1: Rescue of fucosylation in LAD II fibroblasts by complementation with a C. elegans gene.
Figure 2: Sequence comparison of ORF-7 homologs in different species, dendrogram of ORF-7, ORF-11 and other NST family members, and predicted membrane topology of ORF-7.
Figure 3: Subcellular localization of C. elegans and human ORF-7.
Figure 4: Rescue of fucosylation in LAD II fibroblasts by complementation with the human ortholog of ORF-7.
Figure 5: Comparison of a conserved region in the fourth transmembrane domain of ORF-7, which bears the point mutation of LAD II.

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References

  1. Etzioni, A. et al. Recurrent severe infections caused by a novel leukocyte adhesion deficiency. N. Engl. J. Med. 327, 1789–1792 (1992).

    Article  CAS  Google Scholar 

  2. Becker, D.J. & Lowe, J.B. Leukocyte adhesion deficiency type II. Biochim. Biophys. Acta 1455, 193–204 (1999).

    Article  CAS  Google Scholar 

  3. Vestweber, D. & Blanks, J.E. Mechanisms that regulate the function of the selectins and their ligands. Physiol. Rev. 79, 181–213 (1999).

    Article  CAS  Google Scholar 

  4. Marquardt, T. et al. Correction of leukocyte adhesion deficiency type II with oral fucose. Blood 94, 3976–3985 (1999).

    CAS  PubMed  Google Scholar 

  5. Körner, C. et al. Decreased availability of GDP-L-fucose in a patient with LAD II with normal GDP-D-mannose dehydratase and FX protein activities. J. Leukoc. Biol. 66, 95–98 (1999).

    Article  Google Scholar 

  6. Lübke, T., Marquardt, T., von Figura, K. & Körner, C. A new type of carbohydrate-deficient glycoprotein syndrome due to a decreased import of GDP-fucose into the golgi. J. Biol. Chem. 274, 25986–25989 (1999).

    Article  Google Scholar 

  7. Maly, P. et al. The α-(1,3)fucosyltransferase Fuc-TVII controls leukocyte trafficking through an essential role in L-, E-, and P-selectin ligand biosynthesis. Cell 86, 643–653 (1996).

    Article  CAS  Google Scholar 

  8. Marquardt, T. et al. Leukocyte adhesion deficiency II syndrome, a generalized defect in fucose metabolism. J. Pediatr. 134, 681–688 (1999).

    Article  CAS  Google Scholar 

  9. Gerardy-Schahn, R. & Eckhardt, M. Nucleotide sugar transporters in carbohydrates. in Oligosaccharides in Chemistry and Biology: A Comprehensive Handbook, Vol. II. Biology of Saccharides (eds. Ernst, B., Hart, G. & Sinay, P.) 19–36 (Wiley-VCH, Weinheim, 2000).

    Google Scholar 

  10. van Die, I. et al. Core α1,3)-fucose is a common modification of N-glycans in parasitic helminths and constitutes an important epitope for IgE from Haemonchus contortus infected sheep. FEBS Lett. 463, 189–193 (1999).

    Article  CAS  Google Scholar 

  11. DeBose-Boyd, R.A., Nyame, A.K. & Cummings, R.D. Molecular cloning and characterization of an α1,3-fucosyltransferase, CEFT-1, from Caenorhabditis elegans. Glycobiology 8, 905–917 (1998).

    Article  CAS  Google Scholar 

  12. Abeijon, C., Robbins, P. & Hirschberg, C.B. Molecular cloning of the Golgi apparatus uridine diphosphate-N-acetylglucosamine transporter from Kluyveromyces lactis. Proc. Natl. Acad. Sci. USA 93, 5963–5968 (1996).

    Article  CAS  Google Scholar 

  13. Lübke, T. et al. Complementation cloning identifies CDG-IIc, a new type of congenital disorders of glycosylation, as a GDP-fucose transporter deficiency. Nature Genet. 28, 73–76 (2001).

    PubMed  Google Scholar 

  14. Eckhardt, M., Gotza, B. & Gerardy-Schahn, R. Membrane topology of the mammalian CMP-sialic acid transporter. J. Biol. Chem. 274, 8779–8787 (1999).

    Article  CAS  Google Scholar 

  15. Berninsone, P., Hwang, H.-Y., Zemtseva, I., Horvitz, H.R. & Hirschberg, C.B. SQV-7, a protein involved in Caenorhabditis elegans epithelial invagination and early embryogenesis, transports UDP-glucuronic acid, UDP-N-acetylgalactosamine, and UDP-galactose. Proc. Natl. Acad. Sci. USA 98, 3738–3743 (2001).

    Article  CAS  Google Scholar 

  16. Karsan, A. et al. Leukocyte adhesion deficiency type II is a generalized defect of de novo GDP-fucose biosynthesis. Endothelial cell fucosylation is not required for neutrophil rolling on human nonlymphoid endothelium. J. Clin. Invest. 101, 2438–2445 (1998).

    Article  CAS  Google Scholar 

  17. Lühn, K., Marquardt, T., Harms, E. & Vestweber, D. Discontinuation of fucose therapy in LADII causes rapid loss of selectin ligands and rise of leukocyte counts. Blood 97, 330–332 (2001).

    Article  Google Scholar 

  18. The C. elegans Sequencing Consortium. Genome sequence of the nematode C. elegans: a platform for investigating biology. Science 282, 2012–2018 (1998).

  19. Rost, B. PHD: predicting 1D protein structure byprofile based neural networks. Methods Enzymol. 266, 525–539 (1996).

    Article  CAS  Google Scholar 

  20. Kyte, J. & Doolittle, R.F. A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 157, 105–132 (1982).

    Article  CAS  Google Scholar 

  21. Ishida, N. et al. Indispensability of transmembrane domains of Golgi UDP-galactose transporter as revealed by analysis of genetic defects in UDP-galactose transporter-deficient murine Had-1 mutant cell lines and construction of deletion mutants. J. Biochem. (Tokyo) 126, 1107–1117 (1999).

    Article  CAS  Google Scholar 

  22. Gao, X.-D. & Dean, N. Distinct protein domains of the yeast Golgi GDP-mannose transporter mediate oligomer assembly and export from the endoplasmic reticulum. J. Biol. Chem. 275, 17718–17727 (2000).

    Article  CAS  Google Scholar 

  23. Ebnet, K., Schulz, C.U., Meyer-zu-Brickwedde, M.-K., Pendl, G.G. & Vestweber, D. Junctional adhesion molecule (JAM) interacts with the PDZ domain containing proteins AF-6 and ZO-1. J. Biol. Chem. 275, 27979–27988 (2000).

    CAS  PubMed  Google Scholar 

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Acknowledgements

We thank T. Marquardt for the LAD II fibroblasts; M. Aurrand-Lion and B. Imhof for the GFP-JAM cDNA construct; and J. Brosius and S. Stamm for discussions. This work was supported by a grant of the Deutsche Forschungsgemeinschaft, SFB 293 to D.V.

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Author notes

  1. Kerstin Lühn and Martin K. Wild: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut für Zellbiologie, ZMBE, Universität Münster, Münster, and Max-Planck-Institut für Klinische and Physiologische Forschung, Bad Nauheim, Germany

    Kerstin Lühn, Martin K. Wild & Dietmar Vestweber

  2. Institut für Physiologische Chemie, Universität Bonn, Bonn, Germany

    Matthias Eckhardt

  3. Institut für Medizinische Mikrobiologie, Medizinische Hochschule Hannover, Hannover, Germany

    Rita Gerardy-Schahn

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  1. Kerstin Lühn
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  2. Martin K. Wild
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  4. Rita Gerardy-Schahn
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  5. Dietmar Vestweber
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Correspondence to Dietmar Vestweber.

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Lühn, K., Wild, M., Eckhardt, M. et al. The gene defective in leukocyte adhesion deficiency II encodes a putative GDP-fucose transporter. Nat Genet 28, 69–72 (2001). https://doi.org/10.1038/ng0501-69

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