Intermediate filaments control the intracellular distribution of caspases during apoptosis.
Caspases are responsible for a cascade of events controlling the disassembly of apoptotic cells. We now demonstrate that caspase-9 is activated at an early stage of apoptosis in epithelial cells and all its detectable, catalytically active large subunits (both the p35 and p37) are concentrated on cytokeratin fibrils. Immunolabeling of distinctive neoepitopes, exposed by cleavage of procaspase-9 at either Asp315 or Asp330, was co-localized on these fibrils with active caspase-3, caspase-cleaved cytokeratin-18, death-effector-domain containing DNA-binding protein and ubiquitin. Cytokeratin filaments may thus provide a scaffold whereby active subunits of caspase-9 can activate caspase-3 which, in turn, can activate more caspase-9 so forming an amplification loop to facilitate cleavage of cytokeratin-18, disruption of the cytoskeleton and the ensuing formation of cytoplasmic inclusions. These inclusions, formed from the collapse of fibrils, together with their associated components, also contain ubiquitinated proteins, vimentin, heat-shock protein 72, and tumor necrosis factor receptor type-1-associated death domain protein. Many of their constituents, including active caspases, remain sequestered within these inclusions, even after detergent treatment and isolation. Thus, such inclusions do not merely accumulate disrupted cytokeratins but also sequestrate potentially noxious proteins that could injure healthy neighboring cells.
Dinsdale
David
D
Medical Research Council Toxicology Unit, Leicester, United Kingdom. dd5@le.ac.uk
Lee
Justine C
JC
Dewson
Grant
G
Cohen
Gerald M
GM
Peter
Marcus E
ME
D017209:Apoptosis / Q000502:physiology*; D020169:Caspases / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D002479:Inclusion Bodies / Q000648:ultrastructure; D007382:Intermediate Filaments / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D011233:Precipitin Tests; D021381:Protein Transport / Q000502:physiology; D014407:Tumor Cells, Cultured
D020169:Caspases