Structure of the Ets-1 pointed domain and mitogen-activated protein kinase phosphorylation site
| Autor: | Cameron D. Mackereth, Lawrence P. McIntosh, Barbara J. Graves, Lisa Gentile, Jeffrey J. Seidel, Carolyn M. Slupsky, Logan W. Donaldson |
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| Rok vydání: | 1998 |
| Předmět: |
Magnetic Resonance Spectroscopy
Protein Conformation Molecular Sequence Data Biology Protein–protein interaction Proto-Oncogene Protein c-ets-1 Mice Protein structure Proto-Oncogene Proteins Animals Gene family Amino Acid Sequence Phosphorylation Peptide sequence Transcription factor Calcium-Calmodulin-Dependent Protein Kinases Genetics Multidisciplinary Proto-Oncogene Proteins c-ets Sequence Homology Amino Acid Biological Sciences Fusion protein Cell biology Transcription Factors |
| Zdroj: | Proceedings of the National Academy of Sciences. 95:12129-12134 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.95.21.12129 |
| Popis: | The Pointed (PNT) domain and an adjacent mitogen-activated protein (MAP) kinase phosphorylation site are defined by sequence conservation among a subset of ets transcription factors and are implicated in two regulatory strategies, protein interactions and posttranslational modifi- cations, respectively. By using NMR, we have determined the structure of a 110-residue fragment of murine Ets-1 that includes the PNT domain and MAP kinase site. The Ets-1 PNT domain forms a monomeric five-helix bundle. The architec- ture is distinct from that of any known DNA- or protein- binding module, including the helix-loop-helix fold proposed for the PNT domain of the ets protein TEL. The MAP kinase site is in a highly f lexible region of both the unphosphorylated and phosphorylated forms of the Ets-1 fragment. Phosphor- ylation alters neither the structure nor monomeric state of the PNT domain. These results suggest that the Ets-1 PNT domain functions in heterotypic protein interactions and support the possibility that target recognition is coupled to structuring of the MAP kinase site. Transcription factor families are defined by highly conserved DNA-binding domains that display similar DNA recognition properties. The means by which individual family members control different genes therefore must be determined by regulatory mechanisms that enhance the specificity of DNA binding. In the ets gene family, which includes at least 18 members in the human genome, partnerships with additional transcription factors, as well as posttranslational modifications, help dictate specificity for distinct targets (1). These regulatory mechanisms converge on a highly conserved '80-aa region termed the Pointed (PNT) domain (2). The PNT domain occurs in approximately one-third of the ets proteins, including Ets-1, Ets-2, GABPa, and TEL from vertebrates, and PNT-P2 and Yan from Drosophila (Fig. 1). This domain is proposed to mediate protein-protein interac- tions and to be regulated by ras-dependent signaling because of the presence of an adjacent mitogen-activated protein (MAP) kinase phosphorylation site (1). In particular, the PNT domain is implicated in the self-association of chimeric onco- proteins, identified in human leukemias, that result from chromosomal translocations of the gene encoding the ets protein TEL with segments of genes encoding several tyrosine kinases or the acute myeloid leukemia (AML)-1B transcrip- tion factor (3-9). To date sequence conservation has defined the PNT do- main, yet it has not been established that this region is a structural module that acts in a biological context. To create a framework for understanding the role of the PNT domain in the regulation of a variety of ets proteins and in the oncogenic potential of TEL fusion proteins, we have characterized struc- turally a fragment of Ets-1 that includes this domain and the adjacent MAP kinase phosphorylation site. |
| Databáze: | OpenAIRE |
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