Defining the ligand specificity of the deleted in colorectal cancer (DCC) receptor

PLoS One. 2014 Jan 6;9(1):e84823. doi: 10.1371/journal.pone.0084823. eCollection 2014.

Abstract

The growth and guidance of many axons in the developing nervous system require Netrin-mediated activation of Deleted in Colorectal Cancer (DCC) and other still unknown signaling cues. Commissural axon guidance defects are more severe in DCC mutant mice than Netrin-1 mutant mice, suggesting additional DCC activating signals besides Netrin-1 are involved in proper axon growth. Here we report that interaction screens on extracellular protein microarrays representing over 1,000 proteins uniquely identified Cerebellin 4 (CBLN4), a member of the C1q-tumor necrosis factor (TNF) family, and Netrin-1 as extracellular DCC-binding partners. Immunofluorescence and radio-ligand binding studies demonstrate that Netrin-1 competes with CBLN4 binding at an overlapping site within the membrane-proximal fibronectin domains (FN) 4-6 of DCC and binds with ∼5-fold higher affinity. CBLN4 also binds to the DCC homolog, Neogenin-1 (NEO1), but with a lower affinity compared to DCC. CBLN4-null mice did not show a defect in commissural axons of the developing spinal cord but did display a transient increase in the number of wandering axons in the brachial plexus, consistent with a role in axon guidance. Overall, the data solidifies CBLN4 as a bona fide DCC ligand and strengthens its implication in axon guidance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / metabolism
  • Carrier Proteins
  • DCC Receptor
  • Embryonic Development / genetics
  • Humans
  • Kinetics
  • Ligands
  • Mice
  • Mutation
  • Nerve Growth Factors / metabolism
  • Nerve Tissue Proteins / metabolism
  • Netrin-1
  • Neurogenesis / genetics
  • Neurons / metabolism
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Interaction Mapping
  • Protein Precursors / metabolism
  • Receptors, Cell Surface / chemistry
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*
  • Tumor Suppressor Proteins / chemistry
  • Tumor Suppressor Proteins / genetics
  • Tumor Suppressor Proteins / metabolism*

Substances

  • Carrier Proteins
  • DCC Receptor
  • DCC protein, human
  • Ligands
  • NTN1 protein, human
  • Nerve Growth Factors
  • Nerve Tissue Proteins
  • Ntn1 protein, mouse
  • Protein Precursors
  • Receptors, Cell Surface
  • Tumor Suppressor Proteins
  • cerebellin 4 precursor, mouse
  • Netrin-1

Grants and funding

All funding for this research was provided by Genentech, a member of the Roche group. The funder did have roles in study design, data collection and analysis, decision to publish, and preparation of the manuscript.