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Molecular and Cellular Biology, January 2000, p. 196-204, Vol. 20, No. 1
0270-7306/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Role of Apoptosis Signal-Regulating Kinase in Regulation of the c-Jun N-Terminal Kinase Pathway and Apoptosis in Sympathetic Neurons

Takashi Kanamoto,1,2 Monica Mota,3 Kohsuke Takeda,1,4 Lee L. Rubin,5 Kohei Miyazono,1 Hidenori Ichijo,1,4,* and Chantal E. Bazenet3,*

Department of Biochemistry, The Cancer Institute, Japanese Foundation for Cancer Research, 1-37-1 Kami-Ikebukuro, Toshima-ku, Tokyo 170,1 Department of Ophthalmology, Hiroshima University School of Medicine, 1-2-3 Kasumi, Minani-ku, Hiroshima 734,2 and Department of Biomaterials Science, Faculty of Dentistry, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549,4 Japan; EISAI London Research Laboratories, University College London, London WC1E 6BT, United Kingdom3; and Ontogeny Inc., Cambridge, Massachusetts 023195

Received 4 February 1999/Returned for modification 26 March 1999/Accepted 10 September 1999

We have previously shown that nerve growth factor (NGF) withdrawal-induced death requires the activity of the small GTP-binding protein Cdc42 and that overexpression of an active form of Cdc42 is sufficient to mediate neuronal apoptosis via activation of the c-Jun pathway. Recently, a new mitogen-activated protein (MAP) kinase kinase kinase, apoptosis signal-regulating kinase 1 (ASK1) which activates both the c-Jun N-terminal kinase (JNK) and p38 MAP kinase pathways and plays pivotal roles in tumor necrosis factor- and Fas-induced apoptosis, has been identified. Therefore, we investigated the role of ASK1 in neuronal apoptosis by using rat pheochromocytoma (PC12) neuronal cells and primary rat sympathetic neurons (SCGs). Overexpression of ASK1-Delta N, a constitutively active mutant of ASK1, activated JNK and induced apoptosis in differentiated PC12 cells and SCG neurons. Moreover, in differentiated PC12 cells, NGF withdrawal induced a four- to fivefold increase in the activity of endogenous ASK1. Finally, expression of a kinase-inactive ASK1 significantly blocked both NGF withdrawal- and Cdc42-induced death and activation of c-jun. Taken together, these results demonstrate that ASK1 is a crucial element of NGF withdrawal-induced activation of the Cdc42-c-Jun pathway and neuronal apoptosis.

* Corresponding author. Mailing address for Hidenori Ichijo: Department of Biomaterials Science, Faculty of Dentistry, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan. Phone: 81 3 58 03 5671. Fax: 81 3 5803 0192. E-mail: ichijo.det2{at}dent.tmd.ac.jp. Mailing address for Chantal E. Bazenet: EISAI London Research Laboratories, University College London, Gower St., London WC16 6BT, United Kingdom. Phone: 44 171-388 4746. Fax: 44 171-413 1121. E-mail: Chantal_bazenet{at}eisai.net.

Molecular and Cellular Biology, January 2000, p. 196-204, Vol. 20, No. 1
0270-7306/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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