Deciphering the Fringe-Mediated Notch Code: Identification of Activating and Inhibiting Sites Allowing Discrimination between Ligands

Dev Cell. 2017 Jan 23;40(2):193-201. doi: 10.1016/j.devcel.2016.12.013. Epub 2017 Jan 12.

Abstract

Fringe proteins are β3-N-acetylglucosaminyltransferases that modulate Notch activity by modifying O-fucose residues on epidermal growth factor-like (EGF) repeats of Notch. Mammals have three Fringes: Lunatic, Manic, and Radical. While Lunatic and Manic Fringe inhibit Notch1 activation from Jagged1 and enhance activation from Delta-like 1, Radical Fringe enhances signaling from both. We used a mass spectrometry approach to determine whether the variable effects of Fringes on Notch1 result from generation of unique glycosylation patterns on Notch1. We found that Lunatic and Manic Fringe modified similar sites on Notch1, while Radical Fringe modified a subset. Fringe modifications at EGF8 and EGF12 enhanced Notch1 binding to and activation from Delta-like 1, while modifications at EGF6 and EGF36 (added by Manic and Lunatic but not Radical) inhibited Notch1 activation from Jagged1. Combined, these results suggest that Fringe modifications "mark" different regions in the Notch1 extracellular domain for activation or inhibition.

Keywords: EGF repeats; Fringe; Notch; O-fucose; development; glycosylation; signal transduction.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Calcium-Binding Proteins
  • Epidermal Growth Factor / metabolism
  • Fucose / metabolism
  • Glucosyltransferases
  • Glycosyltransferases / metabolism*
  • HEK293 Cells
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Jagged-1 Protein / metabolism*
  • Ligands
  • Mice
  • Models, Biological
  • NIH 3T3 Cells
  • Protein Binding
  • Proteins / metabolism*
  • Receptor, Notch1 / chemistry
  • Receptor, Notch1 / metabolism*
  • Repetitive Sequences, Amino Acid

Substances

  • Calcium-Binding Proteins
  • Dlk1 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • Jagged-1 Protein
  • Ligands
  • Proteins
  • Receptor, Notch1
  • Fucose
  • Epidermal Growth Factor
  • Glycosyltransferases
  • Glucosyltransferases
  • Lfng protein, mouse
  • Mfng protein, mouse
  • Rfng protein, mouse