Rhbdf2 mutations increase its protein stability and drive EGFR hyperactivation through enhanced secretion of amphiregulin

Proc Natl Acad Sci U S A. 2014 May 27;111(21):E2200-9. doi: 10.1073/pnas.1323908111. Epub 2014 May 13.

Abstract

The rhomboid 5 homolog 2 (Rhbdf2) gene encodes an inactive rhomboid (iRhom) protease, iRhom2, one of a family of enzymes containing a long cytosolic N terminus and a dormant peptidase domain of unknown function. iRhom2 has been implicated in epithelial regeneration and cancer growth through constitutive activation of epidermal growth factor receptor (EGFR) signaling. However, little is known about the physiological substrates for iRhom2 or the molecular mechanisms underlying these functions. We show that iRhom2 is a short-lived protein whose stability can be increased by select mutations in the N-terminal domain. In turn, these stable variants function to augment the secretion of EGF family ligands, including amphiregulin, independent of metalloprotease a disintegrin and metalloproteinase 17 (ADAM17) activity. In vivo, N-terminal iRhom2 mutations induce accelerated wound healing as well as accelerated tumorigenesis, but they do not drive spontaneous tumor development. This work underscores the physiological prominence of iRhom2 in controlling EGFR signaling events involved in wound healing and neoplastic growth, and yields insight into the function of key iRhom2 domains.

Keywords: ERAD; curly bare; epithelial cancer; pseudoenzyme; tylosis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amphiregulin
  • Animals
  • COS Cells
  • Carrier Proteins / genetics*
  • Chlorocebus aethiops
  • Cloning, Molecular
  • EGF Family of Proteins
  • Enzyme-Linked Immunosorbent Assay
  • ErbB Receptors / metabolism*
  • Glycoproteins / metabolism*
  • HEK293 Cells
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • Immunoprecipitation
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Kaplan-Meier Estimate
  • Mice
  • Mutagenesis
  • Mutation / genetics
  • Protein Stability*
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics
  • Signal Transduction / physiology*
  • Wound Healing / genetics

Substances

  • AREG protein, human
  • Amphiregulin
  • Areg protein, mouse
  • Carrier Proteins
  • EGF Family of Proteins
  • Glycoproteins
  • Intercellular Signaling Peptides and Proteins
  • iRhom2 protein, mouse
  • ErbB Receptors