| First Author | Burns KA | Year | 2011 |
| Journal | J Biol Chem | Volume | 286 |
| Issue | 14 | Pages | 12640-9 |
| PubMed ID | 21285458 | Mgi Jnum | J:263759 |
| Mgi Id | MGI:6193788 | Doi | 10.1074/jbc.M110.187773 |
| Citation | Burns KA, et al. (2011) Selective mutations in estrogen receptor alpha D-domain alters nuclear translocation and non-estrogen response element gene regulatory mechanisms. J Biol Chem 286(14):12640-9 |
| abstractText | The three main mechanisms of ERalpha action are: 1) nuclear, genomic, direct DNA binding, 2) nuclear, genomic, "tethered"-mediated, protein-protein interactions, and 3) non-nuclear, non-genomic, rapid action responses. Reports suggest the D-domain or hinge region of ERalpha plays an important role in mechanisms 1 and 2 above. Studies demonstrating the functionality of the ERalpha hinge region have resected the full D-domain; therefore, site directed mutations were made to attribute precise sequence functionality to this domain. This study focuses on the characterization and properties of three novel site directed ERalpha- D-domain mutants. The Hinge 1 (H1) ERalpha mutant has disrupted nuclear localization, can no longer perform tethered mediated responses and has lost interaction with c-Jun, but retains estrogen response element (ERE)-mediated functions as demonstrated by confocal microscopy, reporter assays, endogenous gene expression and co-immunoprecipitation. The H2 ERalpha mutant is non-nuclear, but translocates to the nucleus with estradiol (E2) treatment and maintains ERE-mediated functionality. The H2+NES ERalpha mutant does not maintain nuclear translocation with hormone binding, no longer activates ERE-target genes, functions in ERE- or tethered-mediated luciferase assays, but does retain the non-genomic, non-nuclear, rapid action response. These studies reveal the sequence(s) in the ERalpha hinge region that are involved in tethered-mediated actions as well as nuclear localization and attribute important functionality to this region of the receptor. In addition, the properties of these ERalpha mutants will allow future studies to further dissect and characterize the three main ERalpha mechanisms of action and determine the mechanistic role each action has in estrogen hormone regulation. |
