Cyclin D1 Expression Is Regulated Positively by the p42/p44MAPK and Negatively by the p38/HOGMAPK Pathway

doi:10.1074/jbc.271.34.20608

Cyclin D1 Expression Is Regulated Positively by the p42/p44^MAPK and Negatively by the p38/HOG^MAPK Pathway*

From the Centre de Biochimie, CNRS-UMR 134, Faculté des Sciences, Parc Valrose, 06108 Nice Cedex 02, France and the
^§ Institut für Molekularbiologie und Tumorforschung (IMT), Philipps-Universität Marburg, Emil-Mannkopff-Strasse 2, D-35037 Marburg, Federal Republic of Germany

^¶ To whom correspondence and reprint request should be addressed. E-mail: pouyssegur{at}uniee.fr.

↵^‡ Present address: Dept. of Biochemistry, Rm. 906B, McIntyre Medical Sciences Bldg., McGill University, 3655 Drummond St., Montreal H3G 1Y6, Quebec, Canada. Tel.: 514-398-8168; Fax: 514-398-7384. E-mail: Lavoie{at}medcor.mcgill.ca.

Abstract

We have previously shown that the persistent activation of p42/p44^MAPK is required to pass the G₁ restriction point in fibroblasts (Pagès, G., Lenormand, P., L'Allemain, G., Chambard, J. C., Meloche, S., and Pouysségur, J. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 8319-8323) and postulated that MAPKs control the activation of G₁ cyclin-dependent complexes. We examined the mitogen-dependent induction of cyclin D1 expression, one of the earliest cell cycle-related events to occur during the G₀/G₁ to S-phase transition, as a potential target of MAPK regulation. Effects exerted either by the p42/p44^MAPK or the p38/HOG^MAPK cascade on the regulation of cyclin D1 promoter activity or cyclin D1 expression were compared in CCL39 cells, using a co-transfection procedure. We found that inhibition of the p42/p44^MAPK signaling by expression of dominant-negative forms of either mitogen-activated protein kinase kinase 1 (MKK1) or p44^MAPK, or by expression of the MAP kinase phosphatase, MKP-1, strongly inhibited expression of a reporter gene driven by the human cyclin D1 promoter as well as the endogenous cyclin D1 protein. Conversely, activation of this signaling pathway by expression of a constitutively active MKK1 mutant dramatically increased cyclin D1 promoter activity and cyclin D1 protein expression, in a growth factor-independent manner. Moreover, the use of a CCL39-derived cell line that stably expresses an inducible chimera of the estrogen receptor fused to a constitutively active Raf-1 mutant (ΔRaf-1:ER) revealed that in absence of growth factors, activation of the Raf > MKK1 > p42/p44MAPK cascade is sufficient to fully induce cyclin D1. In marked contrast, the p38^MAPK cascade showed an opposite effect on the regulation of cyclin D1 expression. In cells co-expressing high levels of the p38^MAPK kinase (MKK3) together with the p38^MAPK, a significant inhibition of mitogen-induced cyclin D1 expression was observed. Furthermore, inhibition of p38^MAPK activity with the specific inhibitor, SB203580, enhanced cyclin D1 transcription and protein level. Altogether, these results support the notion that MAPK cascades drive specific cell cycle responses to extracellular stimuli, at least in part, through the modulation of cyclin D1 expression and associated cdk activities.

Footnotes

↵* This work was supported by grants from CNRS (Centre National de la Recherche Scientifique), INSERM (Institut National de la Santé et de la Recherche Médicale), from l'Association pour la Recherche contre le Cancer (Grant 1021), and MRC (Medical Research Council of Canada) postdoctoral fellowship (to J. N. L.). 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.
↵¹ The abbreviations used are:

MAP

mitogen-activated protein

MAPK

mitogen-activated protein kinase

DMEM

Dulbecco's modified Eagle's medium

FCS

fetal calf serum

PAGE

polyacrylamide gel electrophoresis

MKK

MAP kinase kinase

MKP-1

MAP kinase phosphatase

IL

interleukin

pRb

Retinoblastoma protein

SAPKs

stress-activated protein kinases

ERK

extracellular regulated kinase

ATF

activating transcription factor.
↵²F. McKenzie, unpublished data.
↵³A. Brunet, unpublished data.
↵⁴Lenormand, P., McMahon, M., and Pouysségur, J. (1996) J. Biol. Chem. 271, 15762-15768.
↵⁵G. L'Allemain, J. N. Lavoie, and J. Pouysségur, unpublished data.
↵⁶A. Brunet, J.-M. Brondello, and J. Pouysségur, unpublished results.
↵⁷J.-M. Brondello, F. McKenzie, and J. Pouysségur, unpublished results.
↵⁸P. Lenormand and J. Pouysségur, unpublished results.
↵⁹A. Brunet, J. N. Lavoie, and J. Pouysségur, unpublished results.
↵¹⁰J.-M. Brondello, A. Brunet, and J. Pouysségur, unpublished results.
↵¹¹J. N. Lavoie, A. Brunet, and J. Pouysségur, unpublished results.
↵¹²A. Brunet and J. Pouysségur, unpublished results.
- Received February 21, 1996.
- Revision received May 31, 1996.