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J. Biol. Chem., Vol. 280, Issue 27, 25687-25696, July 8, 2005

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Analysis of Double-stranded RNA-induced Apoptosis Pathways Using Interferon-response Noninducible Small Interfering RNA Expression Vector Library^*

Sahohime Matsumoto¶, Makoto Miyagishi¶||**, Hideo Akashi¶, Ryozo Nagai, and Kazunari Taira¶

From the Department of Chemistry and Biotechnology, School of Engineering, the University of Tokyo, Hongo, Tokyo 113-8656, the Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Hongo, Tokyo 113-8655, ^¶Gene Function Research Center, National Institute of Advanced Industrial Science and Technology, Central 4, 1-1-1 Higashi, Tsukuba Science City 305-8562, and^** 21st Century Center of Excellence Program, Graduate School of Medicine, the University of Tokyo, Hongo, Tokyo 113-8655, Japan

We have developed an original vector library that allowed us to exploit the phenomenon of RNA interference but also allowed us to avoid the confounding effects of the interferon response. In the present work, we used our library of small interfering RNA expression vectors to examine the genes involved in apoptosis that was induced by double-stranded RNA. To our surprise, screening of our library revealed two novel double-stranded RNA-induced apoptotic pathways, a JNK/SAPK-mediated mitochondrial pathway and an ERK2-related pathway, both of which appeared to be independent of the serine-threonine protein kinase-dependent caspase pathway. We also found that MST2 and protein kinase C both activated the pro-apoptotic signal mediated by ERK2. The results of our screening analysis suggested the utility of large scale screenings with libraries of small interfering RNA expression vectors.

Received for publication, November 11, 2004 , and in revised form, April 13, 2005.

^* This work was supported by various grants from the National Institute of Advanced Industrial Science and Technology (AIST), the New Energy and Industrial Technology Development Organization (NEDO), and the Japan Foundation for Aging and Health. 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.

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^|| To whom correspondence may be addressed. Tel.: 81-29-861-3015; Fax: 81-3-5841-7340; E-mail: makoto-m{at}chembio.t.u-tokyo.ac.jp. To whom correspondence may be addressed. Tel.: 81-3-5841-8828; Fax: 81-3-5841-8828; E-mail: taira{at}chembio.t.u-tokyo.ac.jp.

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