A novel Apaf-1-independent putative caspase-2 activation complex

doi:10.1083/jcb.200209004

Published online 2 December 2002. doi:10.1083/jcb.200209004

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© The Rockefeller University Press, 0021-9525/2002/12/739 $5.00
The Journal of Cell Biology, Volume 159, Number 5, 739-745

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A novel Apaf-1–independent putative caspase-2 activation complex

Stuart H. Read¹, Belinda C. Baliga¹, Paul G. Ekert², David L. Vaux² and Sharad Kumar¹^,3

¹ Hanson Institute, e Road, Adelaide, Australia 5000
² Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia 3050
³ Department of Medicine, Adelaide University, Adelaide, Australia 5005

Address correspondence to Sharad Kumar, Hanson Institute, P.O. Box 14, Rundle Mall, Adelaide, Australia 5000. Tel.: 61-8-8222-3738. Fax: 61-8-8222-3139. E-mail: sharad.kumar{at}imvs.sa.gov.au

CVaspase activation is a key event in apoptosis execution. Instress-induced apoptosis, the mitochondrial pathway of caspaseactivation is believed to be of central importance. In thispathway, cytochrome c released from mitochondria facilitatesthe formation of an Apaf-1 apoptosome that recruits and activatescaspase-9. Recent data indicate that in some cells caspase-9may not be the initiator caspase in stress-mediated apoptosisbecause caspase-2 is required upstream of mitochondria for therelease of cytochrome c and other apoptogenic factors. To determinehow caspase-2 is activated, we have studied the formation ofa complex that mediates caspase-2 activation. Using gel filtrationanalysis of cell lysates, we show that caspase-2 is spontaneouslyrecruited to a large protein complex independent of cytochromec and Apaf-1 and that recruitment of caspase-2 to this complexis sufficient to mediate its activation. Using substrate-bindingassays, we also provide the first evidence that caspase-2 activationmay occur without processing of the precursor molecule. Ourdata are consistent with a model where caspase-2 activationoccurs by oligomerization, independent of the Apaf-1 apoptosome.

Key Words: apoptosis; caspase activation; apoptosome; caspase-9; initiator caspase

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