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

doi:10.1128/MCB.20.1.196-204.2000

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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-bindingprotein Cdc42 and that overexpression of an active form of Cdc42is sufficient to mediate neuronal apoptosis via activation ofthe 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 p38MAP kinase pathways and plays pivotal roles in tumor necrosisfactor- and Fas-induced apoptosis, has been identified. Therefore,we investigated the role of ASK1 in neuronal apoptosis by usingrat pheochromocytoma (PC12) neuronal cells and primary rat sympatheticneurons (SCGs). Overexpression of ASK1- $Delta$ N, a constitutively activemutant of ASK1, activated JNK and induced apoptosis in differentiatedPC12 cells and SCG neurons. Moreover, in differentiated PC12 cells,NGF withdrawal induced a four- to fivefold increase in the activityof endogenous ASK1. Finally, expression of a kinase-inactive ASK1significantly blocked both NGF withdrawal- and Cdc42-induced deathand activation of c-jun. Taken together, these results demonstratethat ASK1 is a crucial element of NGF withdrawal-induced activationof the Cdc42-c-Jun pathway and neuronalapoptosis.

^* Corresponding author. Mailing address for Hidenori Ichijo: Department of Biomaterials Science, Faculty of Dentistry, TokyoMedical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo113-8549, Japan. Phone: 81 3 58 03 5671. Fax: 81 3 5803 0192.E-mail: ichijo.det2{at}dent.tmd.ac.jp. Mailing address for ChantalE. Bazenet: EISAI London Research Laboratories, University CollegeLondon, Gower St., London WC16 6BT, United Kingdom. Phone: 44171-388 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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