Baculovirus inhibitor of apoptosis functions at or upstream of the apoptotic suppressor P35 to prevent programmed cell death.
Members of the inhibitor of apoptosis (iap) gene family prevent programmed cell death induced by multiple signals in diverse organisms, suggesting that they act at a conserved step in the apoptotic pathway. To investigate the molecular mechanism of iap function, we expressed epitope-tagged Op-iap, the prototype viral iap from Orgyia pseudotsugata nuclear polyhedrosis virus, by using novel baculovirus recombinants and stably transfected insect cell lines. Epitope-tagged Op-iap blocked both virus- and UV radiation-induced apoptosis. With or without apoptotic stimuli, Op-IAP protein (31 kDa) cofractionated with cellular membranes and the cytosol, suggesting a cytoplasmic site of action. To identify the step(s) at which Op-iap blocks apoptosis, we monitored the effect of Op-iap expression on in vivo activation of the insect CED-3/ICE death proteases (caspases). Op-iap prevented in vivo caspase-mediated cleavage of the baculovirus substrate inhibitor P35 and blocked caspase activity upon viral infection or UV irradiation. However, unlike the stoichiometric inhibitor P35, Op-IAP failed to affect activated caspase as determined by in vitro protease assays. These findings provide the first biochemical evidence that Op-iap blocks activation of the host caspase or inhibits its activity by a mechanism distinct from P35. Moreover, as suggested by the capacity of Op-iap to block apoptosis induced by diverse signals, including virus infection and UV radiation, iap functions at a central point at or upstream from steps involving the death proteases.
Manji
G A
GA
Department of Biochemistry, Graduate School and College of Agricultural and Life Sciences, University of Wisconsin-Madison, 53706, USA.
Hozak
R R
RR
LaCount
D J
DJ
Friesen
P D
PD
D000818:Animals; D017209:Apoptosis / Q000528:radiation effects; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D051034:Inhibitor of Apoptosis Proteins; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011480:Protease Inhibitors / Q000494:pharmacology; D015398:Signal Transduction; D018411:Spodoptera / Q000166:cytology; D014466:Ultraviolet Rays; D014764:Viral Proteins / Q000378:metabolism* / Q000502:physiology*
D015853:Cysteine Proteinase Inhibitors; D051034:Inhibitor of Apoptosis Proteins; D011480:Protease Inhibitors; D014764:Viral Proteins; C086430:inhibitor of apoptosis, Nucleopolyhedrovirus; D003546:Cysteine Endopeptidases