The execution phase of apoptosis is under the control of members of the inhibitor of apoptosis (IAP) family of zinc finger proteins. Several of these proteins contain a C-terminal RING (really interesting new gene) domain that has been postulated to regulate ubiquitination of themselves or their target proteins, thereby modulating thresholds for apoptosis. We demonstrate that the auto-ubiquitination sites of the X-linked IAP (XIAP) are Lys322 and Lys328, located in the third baculovirus IAP repeat domain of the protein. Modification of these sites to arginine dramatically reduces ubiquitination of XIAP, but has no measurable effect on the ability of ectopically expressed IAP to rescue cells from two independent apoptotic inducers. Our data firmly locate the auto-ubiquitination sites, and raise doubts regarding the importance of this event as a mechanism for regulating the levels of XIAP.
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August 2003
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Research Article|
August 01 2003
Identification of ubiquitination sites on the X-linked inhibitor of apoptosis protein
Hwain SHIN;
Hwain SHIN
∗The Program in Apoptosis and Cell Death Research, Burnham Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, U.S.A.
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Kazuya OKADA;
Kazuya OKADA
∗The Program in Apoptosis and Cell Death Research, Burnham Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, U.S.A.
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John C. WILKINSON;
John C. WILKINSON
†Departments of Pathology and Internal Medicine, University of Michigan, Ann Arbor, MI 48109, U.S.A.
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Katherine M. SOLOMON;
Katherine M. SOLOMON
†Departments of Pathology and Internal Medicine, University of Michigan, Ann Arbor, MI 48109, U.S.A.
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Colin S. DUCKETT;
Colin S. DUCKETT
†Departments of Pathology and Internal Medicine, University of Michigan, Ann Arbor, MI 48109, U.S.A.
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John C. REED;
John C. REED
∗The Program in Apoptosis and Cell Death Research, Burnham Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, U.S.A.
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Guy S. SALVESEN
Guy S. SALVESEN
1
∗The Program in Apoptosis and Cell Death Research, Burnham Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, U.S.A.
1To whom correspondence should be addressed (e-mail gsalvesen@burnham.org).
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Publisher: Portland Press Ltd
Received:
April 22 2003
Revision Received:
May 14 2003
Accepted:
May 15 2003
Accepted Manuscript online:
May 15 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 373 (3): 965–971.
Article history
Received:
April 22 2003
Revision Received:
May 14 2003
Accepted:
May 15 2003
Accepted Manuscript online:
May 15 2003
Citation
Hwain SHIN, Kazuya OKADA, John C. WILKINSON, Katherine M. SOLOMON, Colin S. DUCKETT, John C. REED, Guy S. SALVESEN; Identification of ubiquitination sites on the X-linked inhibitor of apoptosis protein. Biochem J 1 August 2003; 373 (3): 965–971. doi: https://doi.org/10.1042/bj20030583
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