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Genes to Cells (2006) 11, 701-717. doi:10.1111/j.1365-2443.2006.00983.x
© 2006 Blackwell Publishing or its licensors
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The initiator caspase, caspase-10ß, and the BH-3-only molecule, Bid, demonstrate evolutionary conservation in Xenopus of their pro-apoptotic activities in the extrinsic and intrinsic pathways

Katsuya Kominami1, Chiyo Takagi2, Tomoko Kurata2, Atsushi Kitayama2, Masami Nozaki3, Tatsuya Sawasaki4, Keisuke Kuida5, Yaeta Endo4, Noboru Manabe6, Naoto Ueno2 and Kazuhiro Sakamaki1,*

1 Department of Animal Development and Physiology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan
2 Department of Developmental Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan
3 Department of Cell Biology, Research Institute for Microbial Diseases, Osaka University, Suita 565-0871, Japan
4 Cell-free Science and Technology Research Center, Ehime University, Matsuyama 790-8577, Japan
5 Department of Biology, Vertex Pharmaceuticals, Cambridge, MA 02139, USA
6 Research Unit for Animal Life Sciences, Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ibaraki-Iwama 319-0206, Japan

Two major apoptotic signaling pathways have been defined in mammals, the extrinsic pathway, initiated by ligation of death receptors, and the intrinsic pathway, triggered by cytochrome c release from mitochondria. Here, we identified and characterized the Xenopus homologs of caspase-10 (xCaspase-10ß), a novel initiator caspase, and Bid (xBid), a BH3-only molecule of the Bcl-2 family involved in both the extrinsic and intrinsic pathways. Exogenous expression of these molecules induced apoptosis of mammalian cells. By biochemical and cytological analyses, we clarified that xCaspase-10ß and xBid exhibit structural and functional similarities to their mammalian orthologues. We also detected xCaspase-10ß and xBid transcripts during embryogenesis by whole-mount in situ hybridization and RT-PCR analysis. Microinjection of mRNA encoding a protease-defect xCaspase-10ß mutant into embryos resulted in irregular development. Enforced expression of active xBid induced cell death in developing embryos. Using transgenic frogs established to allow monitoring of caspase activation in vivo, we confirmed that this form of cell death is caspase-dependent apoptosis. Thus, we demonstrated that the machinery governing the extrinsic and intrinsic apoptotic pathways are already established in Xenopus embryos. Additionally, we propose that the functions of the initiator caspase and BH3-only molecule are evolutionarily conserved in vertebrates, functioning during embryonic development.

Communicated by: Shigekazu Nagata

* Correspondence: E-mail: ksakamak{at}lif.kyoto-u.ac.jp






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