Annual rept. 1 Jul 2000-30 Jun 2001
DUKE UNIV MEDICAL CENTER DURHAM NC
Pagination or Media Count:
In Drosophila melanogaster, a 65 amino acid protein called Reaper is central regulator of apoptosis. Transcriptional induction of reaper is necessary for programmed cell death in flies and reaper deletion prevents apoptosis. Although no vertebrate homologs have been identified, our lab discovered that recombinant Reaper induces rapid apoptosis when added to Xenopus egg extracts. In the extract, Reaper causes cytochrome c release from the mitochondria into the cytosol where it acts a cofactor for the activation of death proteases,, or caspases. Using a Reaper affinity column, our lab purified a Reaper interacting protein named Scythe. Scythe is highly conserved among vertebrates and necessary for Reaper induced apoptosis in the Xenopus egg extract system. The goal of this proposal was to elucidate the mechanism of ReaperScythe action. Toward this end I established assays to measure apoptosis induced by ectopic Reaper expression in cultured human cells, expressed recombinant Reaper and Scythe in human tissue culture cells, and purified a truncated human Scythe protein produced in bacteria. Research progress with respect to the Statement of Work is detailed below.
- Medicine and Medical Research