This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tan, I.
Right arrow Articles by Leung, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tan, I.
Right arrow Articles by Leung, T.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, April 2001, p. 2767-2778, Vol. 21, No. 8
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.8.2767-2778.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Intermolecular and Intramolecular Interactions Regulate Catalytic Activity of Myotonic Dystrophy Kinase-Related Cdc42-Binding Kinase alpha

Ivan Tan,1 Kah Tong Seow,1 Louis Lim,1,2 and Thomas Leung1,*

Glaxo-IMCB Group, Institute of Molecular & Cell Biology, Singapore 117609, Singapore,1 and Institute of Neurology, University College London, London WC1N 1PJ, United Kingdom2

Received 5 October 2000/Returned for modification 3 November 2000/Accepted 31 January 2001

Myotonic dystrophy kinase-related Cdc42-binding kinase (MRCK) is a Cdc42-binding serine/threonine kinase with multiple functional domains. We had previously shown MRCKalpha to be implicated in Cdc42-mediated peripheral actin formation and neurite outgrowth in HeLa and PC12 cells, respectively. Here we demonstrate that native MRCK exists in high-molecular-weight complexes. We further show that the three independent coiled-coil (CC) domains and the N-terminal region preceding the kinase domain are responsible for intermolecular interactions leading to MRCKalpha multimerization. N terminus-mediated dimerization and consequent transautophosphorylation are critical processes regulating MRCKalpha catalytic activities. A region containing the two distal CC domains (CC2 and CC3; residues 658 to 930) was found to interact intramolecularly with the kinase domain and negatively regulates its activity. Its deletion also resulted in an active kinase, confirming a negative autoregulatory role. We provide evidence that the N terminus-mediated dimerization and activation of MRCK and the negative autoregulatory kinase-distal CC interaction are two mutually exclusive events that tightly regulate the catalytic state of the kinase. Disruption of this interaction by a mutant kinase domain resulted in increased kinase activity. MRCK kinase activity was also elevated when cells were treated with phorbol ester, which can interact directly with a cysteine-rich domain next to the distal CC domain. We therefore suggest that binding of phorbol ester to MRCK releases its autoinhibition, allowing N-terminal dimerization and subsequent kinase activation.

* Corresponding author. Mailing address: Glaxo-IMCB Group, Institute of Molecular & Cell Biology, 30 Medical Dr., Singapore 117609, Singapore. Phone: (65) 874 6167. Fax: (65) 774 0742. E-mail: mcbthoml{at}imcb.nus.edu.sg.

Molecular and Cellular Biology, April 2001, p. 2767-2778, Vol. 21, No. 8
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.8.2767-2778.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Wen, W., Liu, W., Yan, J., Zhang, M. (2008). Structure Basis and Unconventional Lipid Membrane Binding Properties of the PH-C1 Tandem of Rho Kinases. J. Biol. Chem. 283: 26263-26273 [Abstract] [Full Text]  
  • Choi, S. H., Czifra, G., Kedei, N., Lewin, N. E., Lazar, J., Pu, Y., Marquez, V. E., Blumberg, P. M. (2008). Characterization of the Interaction of Phorbol Esters with the C1 Domain of MRCK (Myotonic Dystrophy Kinase-related Cdc42 Binding Kinase) {alpha}/{beta}. J. Biol. Chem. 283: 10543-10549 [Abstract] [Full Text]  
  • Black, S. A. Jr., Trackman, P. C. (2008). Transforming Growth Factor-{beta}1 (TGF{beta}1) Stimulates Connective Tissue Growth Factor (CCN2/CTGF) Expression in Human Gingival Fibroblasts through a RhoA-independent, Rac1/Cdc42-dependent Mechanism: STATINS WITH FORSKOLIN BLOCK TGF{beta}1-INDUCED CCN2/CTGF EXPRESSION. J. Biol. Chem. 283: 10835-10847 [Abstract] [Full Text]  
  • Garcia, P., Ucurum, Z., Bucher, R., Svergun, D. I., Huber, T., Lustig, A., Konarev, P. V., Marino, M., Mayans, O. (2006). Molecular insights into the self-assembly mechanism of dystrophia myotonica kinase. FASEB J. 20: 1142-1151 [Abstract] [Full Text]  
  • Jacobs, M., Hayakawa, K., Swenson, L., Bellon, S., Fleming, M., Taslimi, P., Doran, J. (2006). The Structure of Dimeric ROCK I Reveals the Mechanism for Ligand Selectivity. J. Biol. Chem. 281: 260-268 [Abstract] [Full Text]  
  • Elbert, M., Rossi, G., Brennwald, P. (2005). The Yeast Par-1 Homologs Kin1 and Kin2 Show Genetic and Physical Interactions with Components of the Exocytic Machinery. Mol. Biol. Cell 16: 532-549 [Abstract] [Full Text]  
  • Blumenstein, L., Ahmadian, M. R. (2004). Models of the Cooperative Mechanism for Rho Effector Recognition: IMPLICATIONS FOR RhoA-MEDIATED EFFECTOR ACTIVATION. J. Biol. Chem. 279: 53419-53426 [Abstract] [Full Text]  
  • Nishiyama, T., Kii, I., Kudo, A. (2004). Inactivation of Rho/ROCK Signaling Is Crucial for the Nuclear Accumulation of FKHR and Myoblast Fusion. J. Biol. Chem. 279: 47311-47319 [Abstract] [Full Text]  
  • Ng, Y., Tan, I., Lim, L., Leung, T. (2004). Expression of the Human Myotonic Dystrophy Kinase-related Cdc42-binding Kinase {gamma} Is Regulated by Promoter DNA Methylation and Sp1 Binding. J. Biol. Chem. 279: 34156-34164 [Abstract] [Full Text]  
  • Shimizu, T., Ihara, K., Maesaki, R., Amano, M., Kaibuchi, K., Hakoshima, T. (2003). Parallel Coiled-coil Association of the RhoA-binding Domain in Rho-kinase. J. Biol. Chem. 278: 46046-46051 [Abstract] [Full Text]  
  • Wansink, D. G., van Herpen, R. E. M. A., Coerwinkel-Driessen, M. M., Groenen, P. J. T. A., Hemmings, B. A., Wieringa, B. (2003). Alternative Splicing Controls Myotonic Dystrophy Protein Kinase Structure, Enzymatic Activity, and Subcellular Localization. Mol. Cell. Biol. 23: 5489-5501 [Abstract] [Full Text]  
  • Chen, X.-q., Tan, I., Ng, C. H., Hall, C., Lim, L., Leung, T. (2002). Characterization of RhoA-binding Kinase ROKalpha Implication of the Pleckstrin Homology Domain in ROKalpha Function Using Region-specific Antibodies. J. Biol. Chem. 277: 12680-12688 [Abstract] [Full Text]  
  • Eda, M., Yonemura, S., Kato, T., Watanabe, N., Ishizaki, T., Madaule, P., Narumiya, S. (2002). Rho-dependent transfer of Citron-kinase to the cleavage furrow of dividing cells. J. Cell Sci. 114: 3273-3284 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»