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Published 29 March 2004. doi:10.1083/jcb.200312041
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 164, Number 7, 979-984
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PKC{alpha} mediates TGFß-induced growth inhibition of human keratinocytes via phosphorylation of S100C/A11



Masakiyo Sakaguchi1, Masahiro Miyazaki1, Hiroyuki Sonegawa1, Mariko Kashiwagi2, Motoi Ohba2, Toshio Kuroki3, Masayoshi Namba4, and Nam-ho Huh1

1 Department of Cell Biology, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan
2 Institute of Molecular Oncology, Showa University, Tokyo 142-8555, Japan
3 Gifu University, Gifu 501-1193, Japan
4 Niimi College, Niimi 718-8585, Japan

Address correspondence to Nam-ho Huh, Dept. of Cell Biology, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikatachou, Okayama 700-8558, Japan. Tel.: 81-86-235-7393. Fax: 81-86-235-7400. email: namu{at}md.okayama-u.ac.jp

Growth regulation of epithelial cells is of major concern because most human cancers arise from them. We demonstrated previously a novel signal pathway involving S100C/A11 for high Ca2+-induced growth inhibition of normal human keratinocytes (Sakaguchi, M., M. Miyazaki, M. Takaishi, Y. Sakaguchi, E. Makino, N. Kataoka, H. Yamada, M. Namba, and N.H. Huh. 2003. J. Cell Biol. 163:825–835). This paper addresses a question whether transforming growth factor ß (TGFß) shares the pathway with high Ca2+. On exposure of the cells to TGFß1, S100C/A11 was phosphorylated, bound to nucleolin, and transferred to the nucleus, resulting in induction of p21WAF1/CIP1 and p15INK4B through activation of Sp1. Protein kinase C {alpha} (PKC{alpha}) was shown to phosphorylate 10Thr of S100C/A11, which is a critical event for the signal transduction. The TGFß1-induced growth inhibition was almost completely mitigated when PKC{alpha} activity was blocked or when S100C/A11 was functionally sequestered. These results indicate that, in addition to the well-characterized Smad-mediated pathway, the PKC{alpha}–S100C/A11-mediated pathway is involved in and essential for the growth inhibition of normal human keratinocytes cells by TGFß1.

Key Words: calcium; nucleolin; p21WAF1/CIP1; PKC; S100

The online version of this article contains supplemental material.

Abbreviations used in this paper: NHK, normal human keratinocyte; PEI, polyethylenimine; TGFß, transforming growth factor ß.


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