| Full text | |
| Author(s): |
Migliorini Figueira, Ana Carolina
[1]
;
Lima, Luis Mauricio T. R.
[2]
;
Lima, Leonardo H. F.
[1]
;
Ranzani, Americo T.
[1]
;
Mule, Guilherme dos Santos
[2]
;
Polikarpov, Igor
[1]
Total Authors: 6
|
| Affiliation: | [1] Univ Sao Paulo, Inst Fis Sao Carlos, BR-13560970 Sao Carlos, SP - Brazil
[2] Univ Fed Rio de Janeiro, Fac Farm, BR-21941590 Rio De Janeiro - Brazil
Total Affiliations: 2
|
| Document type: | Journal article |
| Source: | BIOCHEMISTRY; v. 49, n. 5, p. 893-904, 2010. |
| Web of Science Citations: | 10 |
| Abstract | |
To shed more light on the molecular requirements for recognition of thyroid response elements (TRES) by thyroid receptors (TRs), we compared the specific aspects of DNA TRE recognition by different TR constructs. Using fluorescence anisotropy, we performed a detailed and hierarchical study of TR-TRE binding. This wits done by comparing the binding affinities of three different TR constructs for four different TRE DNA elements, including palindromic sequences and direct repeats (F2, PAL, DR-1, and DR-4) as well as their interactions with nonspecific DNA sequences. The effect of MgCl2 on suppressing of nonselective DNA binding to TR was also investigated. Furthermore, we determined the dissociation constants of the hTR beta DBD (DNA binding domain) and hTR beta DBD-LBD (DNA binding and ligand binding domains) for specific TRES. We found that a minimum DNA recognition peptide derived from DBD (H1TR) is sufficient for recognition and interaction with TREs, whereas scrambled DNA sequences were unrecognized. Additionally, we determined that the TR DBD binds to F2, PAL, and DR-4 with high affinity and similar K-d values. The TR DBD-LBD recognizes all the tested TRES but binds preferentially to F2, with even higher affinity. Finally, our results demonstrate the important role played by LBDs in modulating TR-DNA binding. (AU) | |
| FAPESP's process: | 08/00078-1 - Structure and biophysical studies of complexes of nuclear receptors, ligands, DNA responsive elements and corregulators proteins |
| Grantee: | Ana Carolina Migliorini Figueira |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 06/00182-8 - Structural biophysics of nuclear receptors and related proteins |
| Grantee: | Igor Polikarpov |
| Support Opportunities: | Research Projects - Thematic Grants |