| First Author | Jonsen MD | Year | 1996 |
| Journal | Mol Cell Biol | Volume | 16 |
| Issue | 5 | Pages | 2065-73 |
| PubMed ID | 8628272 | Mgi Jnum | J:32531 |
| Mgi Id | MGI:80025 | Doi | 10.1128/mcb.16.5.2065 |
| Citation | Jonsen MD, et al. (1996) Characterization of the cooperative function of inhibitory sequences in Ets-1. Mol Cell Biol 16(5):2065-73 |
| abstractText | DNA binding by the eukaryotic transcription factor Ets-1 is negatively regulated by an intramolecular mechanism. Quantitative binding assays compared the DNA-binding activities of native Ets-1, three deletion mutants, and three tryptic fragments. Ets-1 and activated Ets-1 polypeptides differed in DNA-binding affinity as much as 23-fold. Inhibition was mediated by two regions flanking the minimal DNA-binding domain. Both regions regulated affinity by enhancing dissociation of the protein-DNA complex. Three lines of evidence indicated that inhibition requires cooperative interaction between the two regions: first, the two inhibitory regions acted through a common mechanism; second, neither region functioned independently of the other; finally, mutation of the C-terminal inhibitory region altered the conformation of the N-terminal inhibitory region. In addition, partial proteolysis detected an identical altered conformation in the N-terminal inhibitory region of Ets-1 bound to DNA. This finding suggested that repression is transiently disrupted during DNA binding. These results provide evidence that the two inhibitory regions of Ets-1 are structurally, as well as functionally, coupled. In addition, conformational change is shown to be a critical component of the inhibition mechanism. A cooperative, allosteric model of autoinhibition is described. Autoinhibition of Ets-1 could be relieved by either protein partner(s) or posttranslational modifications. |
