Scythe: a novel reaper-binding apoptotic regulator.
Reaper is a central regulator of apoptosis in Drosophila melanogaster. With no obvious catalytic activity or homology to other known apoptotic regulators, reaper's mechanism of action has been obscure. We recently reported that recombinant Drosophila reaper protein induced rapid mitochondrial cytochrome c release, caspase activation and apoptotic nuclear fragmentation in extracts of Xenopus eggs. We now report the purification of a 150 kDa reaper-interacting protein from Xenopus egg extracts, which we have named Scythe. Scythe is highly conserved among vertebrates and contains a ubiquitin-like domain near its N-terminus. Immunodepletion of Scythe from extracts completely prevented reaper-induced apoptosis without affecting apoptosis triggered by activated caspases. Moreover, a truncated variant of Scythe lacking the N-terminal domain induced apoptosis even in the absence of reaper. These data suggest that Scythe is a novel apoptotic regulator that is an essential component in the pathway of reaper-induced apoptosis.
Thress
K
K
Department of Pharmacology and Cancer Biology, Duke University Medical Center, C366 LSRC, Research Drive, Durham, NC 27710, USA.
Henzel
W
W
Shillinglaw
W
W
Kornbluth
S
S
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002352:Carrier Proteins; D002457:Cell Extracts / Q000737:chemistry; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000378:metabolism; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D019476:Insect Proteins / Q000737:chemistry; D008928:Mitochondria / Q000378:metabolism; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014981:Xenopus; D029867:Xenopus Proteins
C116358:BAG6 protein, Xenopus; D002352:Carrier Proteins; D002457:Cell Extracts; D003574:Cytochrome c Group; D029721:Drosophila Proteins; D019476:Insect Proteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D010455:Peptides; D011994:Recombinant Proteins; D029867:Xenopus Proteins; C086973:rpr protein, Drosophila; D010447:Peptide Hydrolases; C108711:DEVDase