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 Shaywitz, A. J.
Right arrow Articles by Greenberg, M. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shaywitz, A. J.
Right arrow Articles by Greenberg, M. E.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, December 2000, p. 9409-9422, Vol. 20, No. 24
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Magnitude of the CREB-Dependent Transcriptional Response Is Determined by the Strength of the Interaction between the Kinase-Inducible Domain of CREB and the KIX Domain of CREB-Binding Protein

Adam J. Shaywitz,1,2 Simon L. Dove,3 Jon M. Kornhauser,2,4 Ann Hochschild,3,* and Michael E. Greenberg2,4,*

Program in Biological and Biomedical Sciences1 and the Departments of Microbiology and Molecular Genetics3 and Neurobiology,4 Harvard Medical School, and Division of Neuroscience, Children's Hospital,2 Boston, Massachusetts 02115

Received 3 July 2000/Returned for modification 21 August 2000/Accepted 26 September 2000

The activity of the transcription factor CREB is regulated by extracellular stimuli that result in its phosphorylation at a critical serine residue, Ser133. Phosphorylation of Ser133 is believed to promote CREB-dependent transcription by allowing CREB to interact with the transcriptional coactivator CREB-binding protein (CBP). Previous studies have established that the domain encompassing Ser133 on CREB, known as the kinase-inducible domain (KID), interacts specifically with a short domain in CBP termed the KIX domain and that this interaction depends on the phosphorylation of Ser133. In this study, we adapted a recently described Escherichia coli-based two-hybrid system for the examination of phosphorylation-dependent protein-protein interactions, and we used this system to study the kinase-induced interaction between the KID and the KIX domain. We identified residues of the KID and the KIX domain that are critical for their interaction as well as two pairs of oppositely charged residues that apparently interact at the KID-KIX interface. We then isolated a mutant form of the KIX domain that interacts more tightly with wild-type and mutant forms of the KID than does the wild-type KIX domain. We show that in the context of full-length CBP, the corresponding amino acid substitution resulted in an enhanced ability of CBP to stimulate CREB-dependent transcription in mammalian cells. Conversely, an amino acid substitution in the KIX domain that weakens its interaction with the KID resulted in a decreased ability of full-length CBP to stimulate CREB-dependent transcription. These findings demonstrate that the magnitude of CREB-dependent transcription in mammalian cells depends on the strength of the KID-KIX interaction and suggest that the level of transcription induced by coactivator-dependent transcriptional activators can be specified by the strength of the activator-coactivator interaction.

* Corresponding author. Mailing address for M. E. Greenberg: Division of Neuroscience, Children's Hospital, 250 Enders Research Building, 300 Longwood Ave., Boston, MA 02115. Phone: (617) 355-8344. Fax: (617) 738-1542. E-mail: greenberg{at}a1.tch.harvard.edu. Mailing address for Ann Hochschild: Department of Microbiology and Molecular Genetics, Harvard Medical School, Building D1, 200 Longwood Ave., Boston, MA 02115. Phone: (617) 432-1986. Fax: (617) 738-7664. E-mail: ahochschild{at}hms.harvard.edu.

Molecular and Cellular Biology, December 2000, p. 9409-9422, Vol. 20, No. 24
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Sharma, N., Lopez, D. I., Nyborg, J. K. (2007). DNA Binding and Phosphorylation Induce Conformational Alterations in the Kinase-inducible Domain of CREB: IMPLICATIONS FOR THE MECHANISM OF TRANSCRIPTION FUNCTION. J. Biol. Chem. 282: 19872-19883 [Abstract] [Full Text]  
  • Wood, M. A., Attner, M. A., Oliveira, A. M.M., Brindle, P. K., Abel, T. (2006). A transcription factor-binding domain of the coactivator CBP is essential for long-term memory and the expression of specific target genes. Learn. Mem. 13: 609-617 [Abstract] [Full Text]  
  • Hoffmann, A., Barz, T., Spengler, D. (2006). Multitasking C2H2 Zinc Fingers Link Zac DNA Binding to Coordinated Regulation of p300-Histone Acetyltransferase Activity.. Mol. Cell. Biol. 26: 5544-5557 [Abstract] [Full Text]  
  • Horiuchi, J., Jiang, W., Zhou, H., Wu, P., Yin, J. C. P. (2004). Phosphorylation of Conserved Casein Kinase Sites Regulates cAMP-response Element-binding Protein DNA Binding in Drosophila. J. Biol. Chem. 279: 12117-12125 [Abstract] [Full Text]  
  • Lee, S. Y., Aihara, H., Ellenberger, T., Landy, A. (2004). Two structural features of {lambda} integrase that are critical for DNA cleavage by multimers but not by monomers. Proc. Natl. Acad. Sci. USA 101: 2770-2775 [Abstract] [Full Text]  
  • Malek, J. A., Wierzbowski, J. M., Tao, W., Bosak, S. A., Saranga, D. J., Doucette-Stamm, L., Smith, D. R., McEwan, P. J., McKernan, K. J. (2004). Protein interaction mapping on a functional shotgun sequence of Rickettsia sibirica. Nucleic Acids Res 32: 1059-1064 [Abstract] [Full Text]  
  • Lu, Q., Hutchins, A. E., Doyle, C. M., Lundblad, J. R., Kwok, R. P. S. (2003). Acetylation of cAMP-responsive Element-binding Protein (CREB) by CREB-binding Protein Enhances CREB-dependent Transcription. J. Biol. Chem. 278: 15727-15734 [Abstract] [Full Text]  
  • MALEK, J.A., WIERZBOWSKI, J.M., DASCH, G.A., EREMEVA, M.E., MCEWAN, P.J., MCKERNAN, K.J. (2003). Annotation of Novel Proteins Utilizing A Functional Genome Shotgun Coupled with High-Throughput Protein Interaction Mapping. Cold Spring Harb Symp Quant Biol 68: 331-334 [Abstract]  
  • Xie, A.-Y., Bermudez, V. P., Folk, W. R. (2002). Stimulation of DNA Replication from the Polyomavirus Origin by PCAF and GCN5 Acetyltransferases: Acetylation of Large T Antigen. Mol. Cell. Biol. 22: 7907-7918 [Abstract] [Full Text]  
  • Goto, N. K., Zor, T., Martinez-Yamout, M., Dyson, H. J., Wright, P. E. (2002). Cooperativity in Transcription Factor Binding to the Coactivator CREB-binding Protein (CBP). THE MIXED LINEAGE LEUKEMIA PROTEIN (MLL) ACTIVATION DOMAIN BINDS TO AN ALLOSTERIC SITE ON THE KIX DOMAIN. J. Biol. Chem. 277: 43168-43174 [Abstract] [Full Text]  
  • Zor, T., Mayr, B. M., Dyson, H. J., Montminy, M. R., Wright, P. E. (2002). Roles of Phosphorylation and Helix Propensity in the Binding of the KIX Domain of CREB-binding Protein by Constitutive (c-Myb) and Inducible (CREB) Activators. J. Biol. Chem. 277: 42241-42248 [Abstract] [Full Text]  
  • Shaywitz, A. J., Dove, S. L., Greenberg, M. E., Hochschild, A. (2002). Analysis of Phosphorylation-Dependent Protein-Protein Interactions Using a Bacterial Two-Hybrid System. Sci Signal 2002: pl11-pl11 [Abstract] [Full Text]  
  • Boer, A.-K., Drayer, A. L., Rui, H., Vellenga, E. (2002). Prostaglandin-E2 enhances EPO-mediated STAT5 transcriptional activity by serine phosphorylation of CREB. Blood 100: 467-473 [Abstract] [Full Text]  
  • Doi, J., Takemori, H., Lin, X.-z., Horike, N., Katoh, Y., Okamoto, M. (2002). Salt-inducible Kinase Represses cAMP-dependent Protein Kinase-mediated Activation of Human Cholesterol Side Chain Cleavage Cytochrome P450 Promoter through the CREB Basic Leucine Zipper Domain. J. Biol. Chem. 277: 15629-15637 [Abstract] [Full Text]  
  • Yang, T., Davis, R. J., Chow, C.-W. (2001). Requirement of Two NFATc4 Transactivation Domains for CBP Potentiation. J. Biol. Chem. 276: 39569-39576 [Abstract] [Full Text]  
  • West, A. E., Chen, W. G., Dalva, M. B., Dolmetsch, R. E., Kornhauser, J. M., Shaywitz, A. J., Takasu, M. A., Tao, X., Greenberg, M. E. (2001). Calcium regulation of neuronal gene expression. Proc. Natl. Acad. Sci. USA 98: 11024-11031 [Abstract] [Full Text]  
  • Xu, W., Chen, H., Du, K., Asahara, H., Tini, M., Emerson, B. M., Montminy, M., Evans, R. M. (2001). A Transcriptional Switch Mediated by Cofactor Methylation. Science 294: 2507-2511 [Abstract] [Full Text]  
  • Monroy, M. A., Ruhl, D. D., Xu, X., Granner, D. K., Yaciuk, P., Chrivia, J. C. (2001). Regulation of cAMP-responsive Element-binding Protein-mediated Transcription by the SNF2/SWI-related Protein, SRCAP. J. Biol. Chem. 276: 40721-40726 [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»