Nuclear translocation of caspase-3 is dependent on its proteolytic activation and recognition of a substrate-like protein(s).
Caspase-3 is thought to play an important role(s) in the nuclear morphological changes that occur in apoptotic cells and many nuclear substrates for caspase-3 have been identified despite the cytoplasmic localization of procaspase-3. Therefore, whether activated caspase-3 is localized in the nuclei and how active caspase-3 has access to its nuclear targets are important and unresolved questions. Here we confirmed nuclear localizations for both caspase-3-p17 and caspase-3-p12 subunits of active caspase in apoptotic cells using subcellular fractionation analysis. We also prepared polyclonal and monoclonal antibodies specific for active caspase-3 to define the subcellular localization of active caspase-3. Immunocytochemical observations using anti-active caspase-3 antibodies showed nuclear accumulation of active caspase-3 during apoptosis. In addition, caspase-3, but not caspase-7, translocated from the cytoplasm into the nucleus after induction of apoptosis. Mutations at the cleavage site between the p17 and p12 subunits and the substrate recognition site for the P3 amino acid of the DXXD substrate cleavage motif inhibited nuclear translocation of caspase-3, indicating that nuclear transport of active caspase-3 required proteolytic activation and substrate recognition. These results suggest that active caspase-3 is translocated in association with a substrate-like protein(s) from the cytoplasm into the nucleus during progression through apoptosis.
Kamada
Shinji
S
Molecular and Cell Biology Laboratory, The Salk Institute, La Jolla, California 92037, USA. skamada@kobe-u.ac.jp
Kikkawa
Ushio
U
Tsujimoto
Yoshihide
Y
Hunter
Tony
T
D021581:Active Transport, Cell Nucleus; D017209:Apoptosis; D002352:Carrier Proteins / Q000378:metabolism*; D053148:Caspase 3; D020169:Caspases / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000201:enzymology / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D009154:Mutation; D011499:Protein Processing, Post-Translational; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013379:Substrate Specificity
D002352:Carrier Proteins; D021122:Protein Subunits; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases