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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-
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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