Targeted cell killing by reconstituted caspases.
We have developed a two-component system involving reconstituted caspase (recCaspase) for selective and/or conditional ablation of targeted cells. Caspases, the executioners of programmed cell death, are normally synthesized as inactive zymogens and are activated by proteolytic processing of their subunits. We show here, using two different caspases, Caenorhabditis elegans CED-3 and human Caspase-3, that coexpression of the subunits generates constitutively active caspase activity that leads to cell death. This recCaspase activity, however, occurred only when the subunits associated through binding of linked antiparallel leucine-zipper domains. We exploited the dual-component nature of recCaspases by expressing the individual subunits from combinations of promoters either to target selectively the subset of cells for apoptosis or induce cell death in specific cells at specific times during development. The high degree of target specificity and tight regulation of induction of recCaspase would be advantageous in creating animal models that are ablated for specific cells and in other targeted cell killings.
Chelur
Dattananda S
DS
Department of Biological Sciences, Columbia University, 1012 Fairchild Center, New York, NY 10027, USA. tdat@columbia.edu
Chalfie
Martin
M
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins; D053148:Caspase 3; D020169:Caspases / Q000494:pharmacology*; D016923:Cell Death; D006801:Humans; D009474:Neurons / Q000187:drug effects / Q000473:pathology; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D021122:Protein Subunits; D011994:Recombinant Proteins; D013379:Substrate Specificity
D029742:Caenorhabditis elegans Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D053148:Caspase 3; D020169:Caspases; C089223:ced-3 protein, C elegans