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J. Biol. Chem., Vol. 279, Issue 1, 177-187, January 2, 2004

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Molecular Components of a Cell Death Pathway Activated by Endoplasmic Reticulum Stress^*

Rammohan V. Rao, Karen S. Poksay, Susana Castro-Obregon, Birgit Schilling, Richard H. Row, Gabriel del Rio, Bradford W. Gibson, H. Michael Ellerby¶, and Dale E. Bredesen||

From the Buck Institute for Age Research, Novato, California 94945 and the University of California, San Francisco, California 94143

Alterations in Ca²⁺ homeostasis and accumulation of misfolded proteins in the endoplasmic reticulum (ER) cause ER stress that ultimately leads to programmed cell death. Recent studies have shown that ER stress triggers programmed cell death via an alternative intrinsic pathway of apoptosis that, unlike the intrinsic pathway described previously, is independent of Apaf-1 and cytochrome c. In the present work, we have used a set of complementary approaches, including two-dimensional gel electrophoresis coupled with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry and nano-liquid chromatography-electrospray ionization mass spectrometry with tandem mass spectrometry, RNA interference, co-immunoprecipitation, immunodepletion of candidate proteins, and reconstitution studies, to identify mediators of the ER stress-induced cell death pathway. Our data identify two molecules, valosin-containing protein and apoptosis-linked gene-2 (ALG-2), that appear to play a role in mediating ER stress-induced cell death.

Received for publication, April 29, 2003 , and in revised form, October 15, 2003.

^* This work was supported by National Institutes of Health Grants AG12282 (to D. E. B.), NS33376 (to D. E. B. and R. V. R.), and CA84262 (to H. M. E.), Department of Defense Grant DAMD17-98-8613 (to D. E. B.), and by a grant to the Buck Institute from American Bioscience, Inc. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors should be considered as senior authors.

^¶ To whom correspondence may be addressed: The Buck Institute for Age Research, 8001 Redwood Blvd., Novato, CA 94945. Tel.: 415-209-2089; Fax: 415-209-2230; E-mail: mellerby{at}buckinstitute.org. || To whom correspondence may be addressed: The Buck Institute for Age Research, 8001 Redwood Blvd., Novato, CA 94945. Tel.: 415-209-2090; Fax: 415-209-2230; E-mail: dbredesen{at}buckinstitute.org.

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