Intra- and interdimeric caspase-8 self-cleavage controls strength and timing of CD95-induced apoptosis

Sci Signal. 2014 Mar 11;7(316):ra23. doi: 10.1126/scisignal.2004738.

Abstract

Apoptosis in response to the ligand CD95L (also known as Fas ligand) is initiated by caspase-8, which is activated by dimerization and self-cleavage at death-inducing signaling complexes (DISCs). Previous work indicated that the degree of substrate cleavage by caspase-8 determines whether a cell dies or survives in response to a death stimulus. To determine how a death ligand stimulus is effectively translated into caspase-8 activity, we assessed this activity over time in single cells with compartmentalized probes that are cleaved by caspase-8 and used multiscale modeling to simultaneously describe single-cell and population data with an ensemble of single-cell models. We derived and experimentally validated a minimal model in which cleavage of caspase-8 in the enzymatic domain occurs in an interdimeric manner through interaction between DISCs, whereas prodomain cleavage sites are cleaved in an intradimeric manner within DISCs. Modeling indicated that sustained membrane-bound caspase-8 activity is followed by transient cytosolic activity, which can be interpreted as a molecular timer mechanism reflected by a limited lifetime of active caspase-8. The activation of caspase-8 by combined intra- and interdimeric cleavage ensures weak signaling at low concentrations of CD95L and strongly accelerated activation at higher ligand concentrations, thereby contributing to precise control of apoptosis.

Publication types

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

MeSH terms

  • Apoptosis / physiology*
  • Blotting, Western
  • Caspase 8 / chemistry
  • Caspase 8 / metabolism*
  • Computer Simulation
  • Cytosol / metabolism
  • Death Domain Receptor Signaling Adaptor Proteins / metabolism
  • Dimerization
  • Fas Ligand Protein / metabolism*
  • Flow Cytometry
  • HeLa Cells
  • Humans
  • Image Processing, Computer-Assisted
  • Models, Biological*
  • Signal Transduction / physiology*
  • Single-Cell Analysis

Substances

  • Death Domain Receptor Signaling Adaptor Proteins
  • Fas Ligand Protein
  • Caspase 8