This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sharrocks, A D Articles by Shaw, P E Search for Related Content PubMed PubMed Citation Articles by Sharrocks, A D Articles by Shaw, P E

 Previous Article  |  Next Article 

Mol Cell Biol. 1993 January; 13(1): 123-132

Identification of amino acids essential for DNA binding and dimerization in p67SRF: implications for a novel DNA-binding motif.

Spemann Laboratories, Max-Planck-Institut für Immunobiologie, Freiburg, Germany.

ABSTRACT

The serum response factor (p67SRF) binds to a palindromic sequence in the c-fos serum response element (SRE). A second protein, p62TCF binds in conjunction with p67SRF to form a ternary complex, and it is through this complex that growth factor-induced transcriptional activation of c-fos is thought to take place. A 90-amino-acid peptide, coreSRF, is capable for dimerizing, binding DNA, and recruiting p62TCF. By using extensive site-directed mutagenesis we have investigated the role of individual coreSRF amino acids in DNA binding. Mutant phenotypes were defined by gel retardation and cross-linking analyses. Our results have identified residues essential for either DNA binding or dimerization. Three essential basic amino acids whose conservative mutation severely reduced DNA binding were identified. Evidence which is consistent with these residues being on the face of a DNA binding alpha-helix is presented. A phenylalanine residue and a hexameric hydrophobic box are identified as essential for dimerization. The amino acid phasing is consistent with the dimerization interface being presented as a continuous region on a beta-strand. A putative second alpha-helix acts as a linker between these two regions. This study indicates that p67SRF is a member of a protein family which, in common with many DNA binding proteins, utilize an alpha-helix for DNA binding. However, this alpha-helix is contained within a novel domain structure.

This article has been cited by other articles:

• Iyer, D., Chang, D., Marx, J., Wei, L., Olson, E. N., Parmacek, M. S., Balasubramanyam, A., Schwartz, R. J. (2006). Serum response factor MADS box serine -162 phosphorylation switches proliferation and myogenic gene programs. Proc. Natl. Acad. Sci. USA 103: 4516-4521 [Abstract] [Full Text]  
• Kasza, A., O'Donnell, A., Gascoigne, K., Zeef, L. A. H., Hayes, A., Sharrocks, A. D. (2005). The ETS Domain Transcription Factor Elk-1 Regulates the Expression of Its Partner Protein, SRF. J. Biol. Chem. 280: 1149-1155 [Abstract] [Full Text]  
• Vickers, E. R., Kasza, A., Kurnaz, I. A., Seifert, A., Zeef, L. A. H., O'Donnell, A., Hayes, A., Sharrocks, A. D. (2004). Ternary Complex Factor-Serum Response Factor Complex-Regulated Gene Activity Is Required for Cellular Proliferation and Inhibition of Apoptotic Cell Death. Mol. Cell. Biol. 24: 10340-10351 [Abstract] [Full Text]  
• Stinson, J., Inoue, T., Yates, P., Clancy, A., Norton, J. D., Sharrocks, A. D. (2003). Regulation of TCF ETS-domain transcription factors by helix-loop-helix motifs. Nucleic Acids Res 31: 4717-4728 [Abstract] [Full Text]  
• Zhang, X., Chai, J., Azhar, G., Sheridan, P., Borras, A. M., Furr, M. C., Khrapko, K., Lawitts, J., Misra, R. P., Wei, J. Y. (2001). Early Postnatal Cardiac Changes and Premature Death in Transgenic Mice Overexpressing a Mutant Form of Serum Response Factor. J. Biol. Chem. 276: 40033-40040 [Abstract] [Full Text]  
• Roberts, E. C., Deed, R. W., Inoue, T., Norton, J. D., Sharrocks, A. D. (2001). Id Helix-Loop-Helix Proteins Antagonize Pax Transcription Factor Activity by Inhibiting DNA Binding. Mol. Cell. Biol. 21: 524-533 [Abstract] [Full Text]  
• Dalgleish, P., Sharrocks, A. D. (2000). The mechanism of complex formation between Fli-1 and SRF transcription factors. Nucleic Acids Res 28: 560-569 [Abstract] [Full Text]  
• Ling, Y., West, A. G., Roberts, E. C., Lakey, J. H., Sharrocks, A. D. (1998). Interaction of Transcription Factors with Serum Response Factor. IDENTIFICATION OF THE Elk-1 BINDING SURFACE. J. Biol. Chem. 273: 10506-10514 [Abstract] [Full Text]  
• Groisman, R., Masutani, H., Leibovitch, M.-P., Robin, P., Soudant, I., Trouche, D., Harel-Bellan, A. (1996). Physical Interaction between the Mitogen-responsive Serum Response Factor and Myogenic Basic-Helix-Loop-Helix Proteins. J. Biol. Chem. 271: 5258-5264 [Abstract] [Full Text]  
• Whitmarsh, A., Shore, P, Sharrocks, A., Davis, R. (1995). Integration of MAP kinase signal transduction pathways at the serum response element. Science 269: 403-407 [Abstract]  
• Shore, P., Bisset, L., Lakey, J., Waltho, J. P., Virden, R., Sharrocks, A. D. (1995). Characterization of the Elk-1 ETS DNA-binding Domain. J. Biol. Chem. 270: 5805-5811 [Abstract] [Full Text]  
• Zachgo, S, Silva, E., Motte, P, Trobner, W, Saedler, H, Schwarz-Sommer, Z (1995). Functional analysis of the Antirrhinum floral homeotic DEFICIENS gene in vivo and in vitro by using a temperature-sensitive mutant. Development 121: 2861-2875 [Abstract]  
• Greenall, A., Willingham, N., Cheung, E., Boam, D. S., Sharrocks, A. D. (2001). DNA Binding by the ETS-domain Transcription Factor PEA3 Is Regulated by Intramolecular and Intermolecular Protein{middle dot}Protein Interactions. J. Biol. Chem. 276: 16207-16215 [Abstract] [Full Text]