c-IAP1 Is Cleaved by Caspases to Produce a Proapoptotic C-terminal Fragment

Rollie J. Clem, Ting Ting Sheu, Bettina W.M. Richter, Wei Wu He, Nancy A. Thornberry, Colin S. Duckett, J. Marie Hardwick

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Abstract

Although human c-IAP1 and c-IAP2 have been reported to possess antiapoptotic activity against a variety of stimuli in several mammalian cell types, we observed that full-length c-IAP1 and c-IAP2 failed to protect cells from apoptosis induced by Bax overexpression, tumor necrosis factor α treatment or Sindbis virus infection. However, deletion of the C-terminal RING domains of c-IAP1 and c-IAP2 restored antiapoptotic activity, indicating that this region negatively regulates the antiapoptotic function of the N-terminal BIR domain. This finding is consistent with the observation by others that the spacer region and RING domain of c-IAP1 functions as an E3 ligase, promoting autoubiquitination and degradation of c-IAP1. In addition, we found that c- IAP1 is cleaved during apoptosis to 52- and 35-kDa fragments. Both fragments contain the C-terminal end of c-IAP1 including the RING finger. In vitro cleavage of c-IAP1 with apoptotic cell extracts or with purified recombinant caspase-3 produced similar fragments. Furthermore, transfection of cells with the spacer-RING domain alone suppressed the antiapoptotic function of the N-terminal BIR domain of c-IAP1 and induced apoptosis. Optimal death-inducing activity of the spacer-RING required both the spacer region and the zinc-binding RING domain of c-IAP1 but did not require the caspase recruitment domain located within the spacer region. To the contrary, deletion of the caspase recruitment domain increased proapoptotic activity, apparently by stabilizing the C-terminal fragment.

Original languageEnglish (US)
Pages (from-to)7602-7608
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number10
DOIs
StatePublished - Mar 9 2001

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Clem, R. J., Sheu, T. T., Richter, B. W. M., He, W. W., Thornberry, N. A., Duckett, C. S., & Hardwick, J. M. (2001). c-IAP1 Is Cleaved by Caspases to Produce a Proapoptotic C-terminal Fragment. Journal of Biological Chemistry, 276(10), 7602-7608. https://doi.org/10.1074/jbc.M010259200