Fringe is a glycosyltransferase that modifies Notch

Notch receptors function in highly conserved intercellular signalling pathways that direct cell-fate decisions, proliferation and apoptosis in metazoans. Fringe proteins can positively and negatively modulate the ability of Notch ligands to activate the Notch receptor. Here we establish the biochemical mechanism of Fringe action. Drosophila and mammalian Fringe proteins possess a fucose-specific β1,3 N-acetylglucosaminyltransferase activity that initiates elongation of O-linked fucose residues attached to epidermal growth factor-like sequence repeats of Notch. We obtained biological evidence that Fringe-dependent elongation of O-linked fucose on Notch modulates Notch signalling by using co-culture assays in mammalian cells and by expression of an enzymatically inactive Fringe mutant in Drosophila . The post-translational modification of Notch by Fringe represents a striking example of modulation of a signalling event by differential receptor glycosylation and identifies a mechanism that is likely to be relevant to other signalling pathways.

[1]  P. Stanley,et al.  Specific changes in the oligosaccharide moieties of VSV grown in different lectin-resistant CHO cells , 1978, Cell.

[2]  John G Flanagan,et al.  Complementary gradients in expression and binding of ELF-1 and Mek4 in development of the topographic retinotectal projection map , 1995, Cell.

[3]  K. Irvine,et al.  Fringe modulates Notch–ligand interactions , 1997, Nature.

[4]  Reed J. Harris,et al.  O-linked fucose and other post-translational modifications unique to EGF modules. , 1993, Glycobiology.

[5]  J. Boulter,et al.  Jagged: A mammalian ligand that activates notch1 , 1995, Cell.

[6]  S. Munro,et al.  Activity of the yeast MNN1 alpha-1,3-mannosyltransferase requires a motif conserved in many other families of glycosyltransferases. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[7]  K. von Figura,et al.  A New Type of Carbohydrate-deficient Glycoprotein Syndrome Due to a Decreased Import of GDP-fucose into the Golgi* , 1999, The Journal of Biological Chemistry.

[8]  T. Vogt,et al.  Dorsal-ventral signaling in limb development. , 1997, Current opinion in cell biology.

[9]  R. Kelley,et al.  The effect of O-fucosylation on the first EGF-like domain from human blood coagulation factor VII. , 1999, Biochemistry.

[10]  P. Stanley,et al.  Two Chinese hamster ovary glycosylation mutants affected in the conversion of GDP-mannose to GDP-fucose. , 1986, Archives of biochemistry and biophysics.

[11]  P. Robbins,et al.  Transporters of nucleotide sugars, ATP, and nucleotide sulfate in the endoplasmic reticulum and Golgi apparatus. , 1998, Annual review of biochemistry.

[12]  S. Bray,et al.  The Abruptex domain of Notch regulates negative interactions between Notch, its ligands and Fringe. , 2000, Development.

[13]  K. Irvine,et al.  Modulators of Notch signaling. , 1998, Seminars in cell & developmental biology.

[14]  N. Perrimon,et al.  Specificities of heparan sulphate proteoglycans in developmental processes , 2000, Nature.

[15]  S. Kornfeld.  Diseases of abnormal protein glycosylation: an emerging area. , 1998, The Journal of clinical investigation.

[16]  C. Ohyama,et al.  Molecular Cloning and Expression of GDP-d-mannose-4,6-dehydratase, a Key Enzyme for Fucose Metabolism Defective in Lec13 Cells* , 1998, The Journal of Biological Chemistry.

[17]  K D Irvine,et al.  Fringe, Notch, and making developmental boundaries. , 1999, Current opinion in genetics & development.

[18]  R. Kumar,et al.  Molecular Cloning of Human GDP-mannose 4,6-Dehydratase and Reconstitution of GDP-fucose Biosynthesis in Vitro * , 1998, The Journal of Biological Chemistry.

[19]  R. Haltiwanger,et al.  The O-Linked Fucose Glycosylation Pathway , 1997, The Journal of Biological Chemistry.

[20]  R. J. Fleming,et al.  Specific EGF repeats of Notch mediate interactions with Delta and serrate: Implications for notch as a multifunctional receptor , 1991, Cell.

[21]  K. Brew,et al.  Role of a conserved acidic cluster in bovine beta1,4 galactosyltransferase-1 probed by mutagenesis of a bacterially expressed recombinant enzyme. , 1999, Glycobiology.

[22]  J. Hsieh,et al.  Truncated mammalian Notch1 activates CBF1/RBPJk-repressed genes by a mechanism resembling that of Epstein-Barr virus EBNA2 , 1996, Molecular and cellular biology.

[23]  E. Wieschaus,et al.  fringe, a boundary-specific signaling molecule, mediates interactions between dorsal and ventral cells during Drosophila wing development , 1994, Cell.

[24]  C. MacArthur,et al.  FGF-8 isoforms differ in NIH3T3 cell transforming potential. , 1995, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[25]  M. Muskavitch,et al.  Ligand-receptor interactions and trans-endocytosis of Delta, Serrate and Notch: members of the Notch signalling pathway in Drosophila. , 1999, Journal of cell science.

[26]  S. Artavanis-Tsakonas,et al.  Notch Signaling : Cell Fate Control and Signal Integration in Development , 1999 .

[27]  R. J. Fleming,et al.  Serrate-mediated activation of Notch is specifically blocked by the product of the gene fringe in the dorsal compartment of the Drosophila wing imaginal disc. , 1997, Development.

[28]  William J. Ray,et al.  A presenilin-1-dependent γ-secretase-like protease mediates release of Notch intracellular domain , 1999, Nature.

[29]  N. Perrimon,et al.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. , 1993, Development.

[30]  R. Haltiwanger,et al.  Mammalian Notch1 Is Modified with Two Unusual Forms ofO-Linked Glycosylation Found on Epidermal Growth Factor-like Modules* , 2000, The Journal of Biological Chemistry.

[31]  A. M. Arias,et al.  Interactions among Delta, Serrate and Fringe modulate Notch activity during Drosophila wing development. , 1998, Development.

[32]  S. Carroll,et al.  Fringe forms a complex with Notch , 2000, Nature.

[33]  G. Weinmaster,et al.  Fringe differentially modulates Jagged1 and Delta1 signalling through Notch1 and Notch2 , 2000, Nature Cell Biology.

[34]  S. Artavanis-Tsakonas,et al.  Specific truncations of Drosophila Notch define dominant activated and dominant negative forms of the receptor , 1993, Cell.

[35]  P. Stanley,et al.  Lectin-resistant CHO cells: Selection of four new pea lectin-resistant phenotypes , 1986, Somatic cell and molecular genetics.

[36]  P. Bork,et al.  Secreted Fringe-like Signaling Molecules May Be Glycosyltransferases , 1997, Cell.

[37]  S E Lux,et al.  Constitutively active human Notch1 binds to the transcription factor CBF1 and stimulates transcription through a promoter containing a CBF1-responsive element. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[38]  A. Garcı́a-Bellido,et al.  Roles of the Notch gene in Drosophila wing morphogenesis , 1994, Mechanisms of Development.

[39]  S. Pestka,et al.  Human cytomegalovirus harbors its own unique IL-10 homolog (cmvIL-10). , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[40]  S. Carroll,et al.  Cell recognition, signal induction, and symmetrical gene activation at the dorsal-ventral boundary of the developing drosophila wing , 1995, Cell.

[41]  Raphael Kopan,et al.  A ligand-induced extracellular cleavage regulates gamma-secretase-like proteolytic activation of Notch1. , 2000, Molecular cell.

[42]  A. Joutel,et al.  Notch signalling pathway and human diseases. , 1998, Seminars in cell & developmental biology.

[43]  M. W. Young,et al.  Mutations altering the structure of epidermal growth factor-like coding sequences at the Drosophila Notch locus , 1987, Cell.

[44]  K D Irvine,et al.  A family of mammalian Fringe genes implicated in boundary determination and the Notch pathway. , 1997, Development.

[45]  R. Hill,et al.  Glycosyltransferases and their use in assessing oligosaccharide structure and structure-function relationships. , 1981, Advances in enzymology and related areas of molecular biology.

[46]  P. Stanley,et al.  Chinese hamster ovary cells selected for resistance to the cytotoxicity of phytohemagglutinin are deficient in a UDP-N-acetylglucosamine--glycoprotein N-acetylglucosaminyltransferase activity. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[47]  D. Zinyk,et al.  Fringe boundaries coincide with Notch-dependent patterning centres in mammals and alter Notch-dependent development in Drosophila , 1997, Nature Genetics.

[48]  A Cumano,et al.  A novel proteolytic cleavage involved in Notch signaling: the role of the disintegrin-metalloprotease TACE. , 2000, Molecular cell.

[49]  R. Haltiwanger,et al.  The O-linked fucose glycosylation pathway: identification and characterization of a uridine diphosphoglucose: fucose-beta1,3-glucosyltransferase activity from Chinese hamster ovary cells. , 1999, Glycobiology.