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

Research output: Contribution to journalArticle

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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Caspases
Apoptosis
Cells
Sindbis Virus
Ubiquitin-Protein Ligases
Virus Diseases
Cell Extracts
Viruses
Caspase 3
Transfection
Zinc
Tumor Necrosis Factor-alpha
Observation
Degradation
Caspase Activation and Recruitment Domain

ASJC Scopus subject areas

  • Biochemistry

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

c-IAP1 Is Cleaved by Caspases to Produce a Proapoptotic C-terminal Fragment. / Clem, Rollie J.; Sheu, Ting Ting; Richter, Bettina W M; He, Wei Wu; Thornberry, Nancy A.; Duckett, Colin S.; Hardwick, J Marie.

In: Journal of Biological Chemistry, Vol. 276, No. 10, 09.03.2001, p. 7602-7608.

Research output: Contribution to journalArticle

Clem, RJ, Sheu, TT, Richter, BWM, He, WW, Thornberry, NA, Duckett, CS & Hardwick, JM 2001, 'c-IAP1 Is Cleaved by Caspases to Produce a Proapoptotic C-terminal Fragment', Journal of Biological Chemistry, vol. 276, no. 10, pp. 7602-7608. https://doi.org/10.1074/jbc.M010259200
Clem RJ, Sheu TT, Richter BWM, He WW, Thornberry NA, Duckett CS et al. c-IAP1 Is Cleaved by Caspases to Produce a Proapoptotic C-terminal Fragment. Journal of Biological Chemistry. 2001 Mar 9;276(10):7602-7608. https://doi.org/10.1074/jbc.M010259200
Clem, Rollie J. ; Sheu, Ting Ting ; Richter, Bettina W M ; He, Wei Wu ; Thornberry, Nancy A. ; Duckett, Colin S. ; Hardwick, J Marie. / c-IAP1 Is Cleaved by Caspases to Produce a Proapoptotic C-terminal Fragment. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 10. pp. 7602-7608.
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