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Fringe is a glycosyltransferase that modifies Notch

Nature volume 406, pages 369–375 (2000)Cite this article

Abstract

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.

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Figure 1: Manic Fringe elongates the O-linked fucose saccharides on full-length Notch1 and Notch1 EGF repeats 19–23.
Figure 2: Lunatic, Manic and Drosophila Fringe catalyse the transfer of GlcNAc from UDP-[3H]GlcNAc to fucose in vitro.
Figure 3: Lunatic, Manic and Drosophila Fringe are fucose-specific β1,3 GlcNAc-transferases.
Figure 4: O-linked fucose is required for Jagged1-dependent activation of Notch and Fringe inhibition of CBF1 activation.
Figure 5: Mutation of a conserved acidic patch in Fringe renders it biologically inactive.
Figure 6: Fringe modulates Notch signalling by elongation of O-linked fucose on Notch EGF repeats.

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Acknowledgements

We thank K. Matta for disaccharide standards; K. Severinov for advice on protein purification; T. Correia, S. Narula and S. Sundaram for technical assistance; the Developmental Studies Hybridoma Bank; S. Lux and S. Kotenko for antibodies; G. Weinmaster for Jagged1-expressing mouse L cells and co-culture reagents; J. Flanagan for the AP-Tag plasmid; D. Hayward for CBF1-luciferase reporter plasmid JH26; J. Nye for the mouse Notch1 plasmid; S. Kotenko for Flag expression plasmid; and J. Botas and the Bloomington Stock Center for Drosophila stocks. We thank our colleagues for comments on the manuscript. S.H.J. was supported as a Harold W. Dodds Fellow of the Princeton University Graduate Program. Research in K.D.I.'s laboratory was supported by a GM NIH grant, and by Charles and Joanna Busch fellowships to V.P. and R.W. Research in R.S.H.'s laboratory was supported by a Grant Award from Neose Technologies, Inc. Research in P.S.'s laboratory was supported by a NIH/NCI grant and a NCI Comprehensive Cancer Center grant. Research in T.F.V's lab was supported by an NIH grant and a gift to the Department of Molecular Biology by the Rathmann Family Foundation.

Author information

Author notes

  1. Stuart H. Johnston

    Present address: Exelixis, Inc., South San Francisco, CA, 94080, USA

  2. Thomas F. Vogt

    Present address: Merck Research Laboratories, West Point, PA, 19486-0004, USA

  3. Daniel J. Moloney, Vladislav M. Panin, Stuart H. Johnston, Jihua Chen and Pamela Stanley: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biochemistry and Cell Biology, Institute for Cell and Developmental Biology, SUNY-Stony Brook, Stony Brook, 11794-5215 , New York, USA

    Daniel J. Moloney, Li Shao & Robert S. Haltiwanger

  2. Waksman Institute and Department of Molecular Biology and Biochemistry, Rutgers, The State University, Piscataway, 08854, New Jersey, USA

    Vladislav M. Panin, Richa Wilson & Kenneth D. Irvine

  3. Department of Molecular Biology Princeton University, Princeton, 08544, New Jersey, USA

    Stuart H. Johnston & Thomas F. Vogt

  4. Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, New York, 10461, New York, USA

    Jihua Chen & Pamela Stanley

  5. Department of Pharmacokinetics and Metabolism, Genentech, Inc., South San Francisco, 94080, California , USA

    Yang Wang

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Correspondence to Pamela Stanley, Kenneth D. Irvine, Robert S. Haltiwanger or Thomas F. Vogt.

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Moloney, D., Panin, V., Johnston, S. et al. Fringe is a glycosyltransferase that modifies Notch. Nature 406, 369–375 (2000). https://doi.org/10.1038/35019000

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