Protein Kinase D Induces Transcription through Direct Phosphorylation of the cAMP-response Element-binding Protein*
- Mona Johannessen‡1,
- Marit Pedersen Delghandi‡1,
- An Rykx§,
- Marte Dragset‡,
- Jackie R. Vandenheede§,
- Johan Van Lint§ and
- Ugo Moens‡2
- ‡Department of Microbiology and Virology, Institute of Medical Biology, University of Tromsø, N-9037 Tromsø, Norway and §Molecular Medicine of Protein Kinases, Department of Molecular Cell Biology, Faculty of Medicine, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium
- 2 To whom correspondence should be addressed. Tel.: 47-77-64-46-22; Fax: 47-77-64-53-50; E-mail: ugom{at}fagmed.uit.no.
Abstract
Protein kinase D (PKD), a family of serine/threonine kinases, can be activated by a multitude of stimuli in a protein kinase C-dependent or -independent manner. PKD is involved in signal transduction pathways controlling cell proliferation, apoptosis, motility, and protein trafficking. Despite its versatile functions, few genuine in vivo substrates for PKD have been identified. In this study we demonstrate that the transcription factor cAMP-response element-binding protein (CREB) is a direct substrate for PKD. PKD1 and CREB interact in cells, and activated PKD1 provokes CREB phosphorylation at Ser-133 both in vitro and in vivo. A constitutive active mutant of PKD1 stimulates GAL4-CREB-mediated transcription in a Ser-133-dependent manner, activates CRE-responsive promoters, and increases the expression of CREB target genes. PKD1 also enhances transcription mediated by two other members of the CREB family, ATF-1 and CREM. Our results describe a novel mechanism for PKD-induced signaling through activation of the transcription factor CREB and suggest that stimulus-induced phosphorylation of CREB, reported to be mediated by protein kinase C, may involve downstream activated PKD.
Footnotes
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↵3 The abbreviations used are: PKD, protein kinase D; ca, constitutive active; CREB, cAMP-response element-binding protein; DAG, diacylglycerol; dn, dominant negative; PDGF, platelet-derived growth factor; PH, pleckstrin homology; RT, reverse transcriptase; TPA, 12-O-tetradecanoyl-phorbol-13-acetate; PKC, protein kinase C; GST, glutathione S-transferase; PH, pleckstrin homology; siRNA, small interfering RNA; EGFP, enhanced green fluorescent protein; PKA, protein kinase A; HDAC, histone deacetylase; PBS, phosphate-buffered saline; MBP, myelin basic protein; CREM, cAMP-response element modulator; CRE, cAMP-responsive element; APRT, adenine phosphoribosyltransferase.
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↵4 M. Johannessen, M. P. Delghandi, A. Rykx, M. Dragset, J. R. Vandenheede, J. Van Lint, and U. Moens, unpublished results.
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↵* Work in the Moens laboratory was supported by grants from the Norwegian Research Council, NFR Projects S5168 and S5228, the Norwegian Cancer Society, Kreftforeningen, Project A01037/004, the Aakre Foundation Grant A5048, and The Mjåland Foundation. Work in J. Van Lint and J. R. Vandenheee groups was sponsored by the Belgian Federal Government Grant IAP 5/12, the Belgian Federation against Cancer Grant SCIE2003-53, and by the FWO-Vlaanderen Grants G.0152.03, 1.5.029.04, and G.0282.05. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement”in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵1 Both authors contributed equally to this work.
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- Received November 17, 2006.
- Revision received March 27, 2007.
