Specific Activation of the p38 Mitogen-activated Protein Kinase Signaling Pathway and Induction of Neurite Outgrowth in PC12 Cells by Bone Morphogenetic Protein-2

doi:10.1074/jbc.274.37.26503

Specific Activation of the p38 Mitogen-activated Protein Kinase Signaling Pathway and Induction of Neurite Outgrowth in PC12 Cells by Bone Morphogenetic Protein-2*

From the Laboratory of Cell Regulation, School of Pharmaceutical Sciences, Nagasaki University, 1-14, Bunkyo-machi, Nagasaki 852-8131, Japan and the ^§Laboratory of Cellular Biochemistry, The Institute of Physical and Chemical Research (RIKEN), Wako, Saitama 351-0198, Japan

Abstract

Bone morphogenetic protein (BMP)-2 has the capacity to induce the neuronal differentiation of PC12 cells. Unlike nerve growth factor, however, BMP-2 failed to induce the activation of the 41-/43-kDa mitogen-activated protein (MAP) kinase pathway in these cells. In contrast, BMP-2 characteristically induced the sustained activation of the p38 MAP kinase pathway. Pretreatment of PC12 cells with SB203580 inhibited the BMP-2-induced neurite outgrowth formation in a dose-dependent manner; this inhibition coincided well with the ability of SB203580 to inihibit the BMP-2-induced activation of the p38 MAP kinase pathway. Overexpression in PC12 cells of wild-type MAP kinase kinase (MKK)-6 enhanced the BMP-2-induced activation of p38 MAP kinase, whose activation correlated well with the ability of these cells to induce neurite outgrowth in response to BMP-2. Transient expression of kinase-negative forms of MKK3/6 inhibited the formation of neurite outgrowth in response to BMP-2. Furthermore, expression of constitutively active forms of MKK3/6 induced neurite outgrowth without BMP-2 stimulation, and SB203580 inhibited this induction. These results clearly indicate that activation of the p38 MAP kinase pathway is necessary for BMP-2-induced neuronal differentiation of PC12 cells. Our results also suggest that activation of the p38 MAP kinase pathway alone can induce the neuronal differentiation of PC12 cells.

Footnotes

↵* This work was supported in part by Grants-in-aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan, and a grant from the Naito Foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵‡ Recipient of a fellowship from the Japan Society for the Promotion of Science for Japanese Junior Scientists.
↵¶ To whom all correspondence should be addressed: Laboratory of Cell Regulation, School of Pharmaceutical Sciences, Nagasaki University, 1-14, Bunkyo-machi, Nagasaki 852-8131, Japan. Fax: 81-95-849-2671; E-mail, kohnom@net.nagasaki-u.ac.jp.
↵2 K. Sugiyama and M. Hibi, unpublished data.
↵3 M. Iguchi, K. Watanabe, S. Iwasaki, and M. Kohno, manuscript in preparation.
Abbreviations:

BMP

bone morphogenetic protein

NGF

nerve growth factor

MAP

mitogen-activated protein

ERK

extracellular signal-regulated kinase

MEK

MAP kinase/ERK kinase

JNK

c-Jun N-terminal kinase

MKK

MAP kinase kinase

MAPKAP kinase

MAP kinase-activated protein kinase

FITC

fluorescein isothiocyanate

GST

glutathione S-transferase

HA

hemagglutinin
- Received January 26, 1999.
- Revision received May 18, 1999.
The American Society for Biochemistry and Molecular Biology, Inc.