Regulation of apoptotic signaling pathways in cell-free extracts
ApoptOBis is a program of cellular suicide which rids multicellular otanisms of damaged or extraneous cells. In order to elucidate the cellular signaling pathways which contribute to apoptotic cell death, we are utilizing an extract of Xenopus eggs which can recapitulate the apoptotic process in vitro. Using these extracts, we have found that the SH2/SH3-containlng adaptor protein, crk, is required for in vitro apoptosls. Moreover, proteins interacting with both the SH2 and SH3 domains are critical for transmitting the apoptotic signal. We are currently purifying proteins interacting with these domains. We have also found that the reaper protein of Drosophila Melanogaster, critical for apoptosis in that organism, can induce rapid apoptosis in extracts. Using reaper as a bait in affinity Chromatography experiments, we have purified a reaper-Interacting protein from egg extracts. Characterization of this protein will be discussed.
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