X-linked and cellular IAPs modulate the stability of C-RAF kinase and cell motility

Nat Cell Biol. 2008 Dec;10(12):1447-55. doi: 10.1038/ncb1804. Epub 2008 Nov 16.

Abstract

Inhibitor of apoptosis proteins (IAP) are evolutionarily conserved anti-apoptotic regulators. C-RAF protein kinase is a direct RAS effector protein, which initiates the classical mitogen-activated protein kinase (MAPK) cascade. This signalling cascade mediates diverse biological functions, such as cell growth, proliferation, migration, differentiation and survival. Here we demonstrate that XIAP and c-IAPs bind directly to C-RAF kinase and that siRNA-mediated silencing of XIAP and c-IAPs leads to stabilization of C-RAF in human cells. XIAP binds strongly to C-RAF and promotes the ubiquitylation of C-RAF in vivo through the Hsp90-mediated quality control system, independently of its E3 ligase activity. In addition, XIAP or c-IAP-1/2 knockdown cells showed enhanced cell migration in a C-RAF-dependent manner. XIAP promotes binding of CHIP (carboxy terminal Hsc70-interacting protein), a chaperone-associated ubiquitin ligase, to the C-RAF-Hsp90 complex in vivo. Interfering with CHIP expression resulted in stabilization of C-RAF and enhanced cell migration, as observed in XIAP knockdown cells. Our data show an unexpected role of XIAP and c-IAPs in the turnover of C-RAF protein, thereby modulating the MAPK signalling pathway and cell migration.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Movement*
  • Enzyme Stability
  • Gene Silencing
  • HSP90 Heat-Shock Proteins / metabolism
  • HeLa Cells
  • Humans
  • Protein Binding
  • Proto-Oncogene Proteins c-raf / metabolism*
  • Transfection
  • Ubiquitin-Protein Ligases
  • X-Linked Inhibitor of Apoptosis Protein / metabolism*

Substances

  • HSP90 Heat-Shock Proteins
  • X-Linked Inhibitor of Apoptosis Protein
  • Ubiquitin-Protein Ligases
  • Proto-Oncogene Proteins c-raf