HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 282, Issue 9, 6733-6742, March 2, 2007

This Article Full Text Full Text (PDF) All Versions of this Article: 282/9/6733    most recent M608086200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kunkel, M. T. Articles by Newton, A. C. Search for Related Content PubMed PubMed Citation Articles by Kunkel, M. T. Articles by Newton, A. C. Social Bookmarking                      What's this?

Calcium-dependent Regulation of Protein Kinase D Revealed by a Genetically Encoded Kinase Activity Reporter^*

Maya T. Kunkel, Alex Toker, Roger Y. Tsien^¶, and Alexandra C. Newton¹

From the Department of Pharmacology and ^¶Howard Hughes Medical Institute, University of California at San Diego, La Jolla, California 92093 and the Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215

Protein kinase D (PKD) regulates many diverse cellular functions in response to diacylglycerol. To monitor PKD signaling in live cells, we generated a genetically encoded fluorescent reporter for PKD activity, DKAR (D kinase activity reporter). DKAR expressed in mammalian cells undergoes reversible fluorescence resonance energy transfer changes upon activation and inhibition of endogenous PKD. Surprisingly, we find that agonist-evoked activation of PKD is driven not only by diacylglycerol production, but by Ca²⁺. Furthermore, elevation of intracellular Ca²⁺, in the absence of any other stimulus, is sufficient to activate PKD. Concurrent imaging of Ca²⁺, diacylglycerol, and PKD activity reveals that thapsigargin-mediated elevation of intracellular Ca²⁺ is closely followed by a robust increase in diacylglycerol production, in turn followed by PKD activation. The Ca²⁺-induced production of diacylglycerol and accompanying PKD activation is dependent on phospholipase C activity. These data reveal that Ca²⁺ is a major contributor to the initiation of PKD signaling through positive feedback regulation of diacylglycerol production, unveiling a new mechanism in PKD activation.

Received for publication, August 23, 2006 , and in revised form, December 21, 2006.

^* This work was supported by National Institutes of Health P01 Grants DK54441 (to A. C. N.), CA075134 (to A. T.), and DK07233 (to M. T. K.). 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.

¹ To whom correspondence should be addressed: Dept. of Pharmacology, University of California at San Diego, Leichtag 282, 9500 Gilman Dr., La Jolla, CA 92093-0721. Tel.: 858-534-4527; Fax: 858-822-5888; E-mail: anewton{at}ucsd.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				Y. Liu, M. Contreras, T. Shen, W. R. Randall, and M. F. Schneider {alpha}-Adrenergic signalling activates protein kinase D and causes nuclear efflux of the transcriptional repressor HDAC5 in cultured adult mouse soleus skeletal muscle fibres J. Physiol., March 1, 2009; 587(5): 1101 - 1115. [Abstract] [Full Text] [PDF]

				L. A. Chen, J. Li, S. R. Silva, L. N. Jackson, Y. Zhou, H. Watanabe, K. L. Ives, M. R. Hellmich, and B. M. Evers PKD3 Is the Predominant Protein Kinase D Isoform in Mouse Exocrine Pancreas and Promotes Hormone-induced Amylase Secretion J. Biol. Chem., January 23, 2009; 284(4): 2459 - 2471. [Abstract] [Full Text] [PDF]

				N. Gaudreault, R. M. Perrin, M. Guo, C. P. Clanton, M. H. Wu, and S. Y. Yuan Counter Regulatory Effects of PKC{beta}II and PKC{delta} on Coronary Endothelial Permeability Arterioscler. Thromb. Vasc. Biol., August 1, 2008; 28(8): 1527 - 1533. [Abstract] [Full Text] [PDF]

				J. Bossuyt, K. Helmstadter, X. Wu, H. Clements-Jewery, R. S. Haworth, M. Avkiran, J. L. Martin, S. M. Pogwizd, and D. M. Bers Ca2+/Calmodulin-Dependent Protein Kinase II{delta} and Protein Kinase D Overexpression Reinforce the Histone Deacetylase 5 Redistribution in Heart Failure Circ. Res., March 28, 2008; 102(6): 695 - 702. [Abstract] [Full Text] [PDF]

				M. Avkiran, A. J. Rowland, F. Cuello, and R. S. Haworth Protein Kinase D in the Cardiovascular System: Emerging Roles in Health and Disease Circ. Res., February 1, 2008; 102(2): 157 - 163. [Abstract] [Full Text] [PDF]