Human Fas-associated factor 1 interacts with heat shock protein 70 and negatively regulates chaperone activity

J Biol Chem. 2005 Mar 4;280(9):8125-33. doi: 10.1074/jbc.M406297200. Epub 2004 Dec 13.

Abstract

We examined the cell death-inducing property of human Fas-associated factor 1 (hFAF1) in the heat shock signaling pathway. By employing co-immunoprecipitation and peptide mass fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry, we found that hFAF1 binds to the 70-kDa heat shock protein family (Hsc70/Hsp70). Interaction mapping indicated that the 82-180 sequence of hFAF1 directly binds to the N-terminal region containing sequence 1-120 of Hsc70/Hsp70. This binding is very tight regardless of ATP and heat shock treatment. Hsc70/Hsp70 and hFAF1 co-localized in the cytosol and nucleus and concentrated to the perinuclear region by heat shock treatment. We examined how hFAF1 regulates Hsp70 function, and found that hFAF1 inhibited the Hsp70 chaperone activity of refolding denatured protein substrates, accelerated heat shock-induced SAPK/JNK activation, and raised heat shock-induced cell death in a binding dependent manner. These results suggest that hFAF1 prevents cells from recovery after stress by binding to and inhibiting the chaperone activity of Hsp70.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Adenosine Triphosphate / chemistry
  • Apoptosis Regulatory Proteins
  • Carrier Proteins / metabolism*
  • Cell Line
  • Cell Nucleus / metabolism
  • Cytosol / metabolism
  • Electrophoresis, Gel, Two-Dimensional
  • Gene Expression Regulation*
  • Glutathione Transferase / metabolism
  • HSC70 Heat-Shock Proteins
  • HSP70 Heat-Shock Proteins / chemistry
  • HSP70 Heat-Shock Proteins / metabolism*
  • HeLa Cells
  • Heat-Shock Proteins / metabolism
  • Humans
  • Immunoprecipitation
  • Luciferases / metabolism
  • Mass Spectrometry
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Mitogen-Activated Protein Kinase 8 / metabolism
  • Models, Genetic
  • Molecular Chaperones / metabolism*
  • Plasmids / metabolism
  • Protein Binding
  • Protein Folding
  • Protein Structure, Tertiary
  • Signal Transduction
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Time Factors
  • Transfection

Substances

  • Adaptor Proteins, Signal Transducing
  • Apoptosis Regulatory Proteins
  • Carrier Proteins
  • FAF1 protein, human
  • HSC70 Heat-Shock Proteins
  • HSP70 Heat-Shock Proteins
  • HSPA8 protein, human
  • Heat-Shock Proteins
  • Molecular Chaperones
  • Adenosine Triphosphate
  • Luciferases
  • Glutathione Transferase
  • Mitogen-Activated Protein Kinase 8