Insights on glucocorticoid receptor activity modulation through the binding of rigid steroids

Autores
<div class="autor_fcen" id="6922">Presman, D.M.</div>; <div class="autor_fcen" id="223">Alvarez, L.D.</div>; <div class="autor_fcen" id="4951">Levi, V.</div>; Eduardo, S.; Digman, M.A.; <div class="autor_fcen" id="5439">Martí, M.A.</div>; <div class="autor_fcen" id="8885">Veleiro, A.S.</div>; <div class="autor_fcen" id="1252">Burton, G.</div>; <div class="autor_fcen" id="6520">Pecci, A.</div>
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
Versión publicada
Descripción
Background: The glucocorticoid receptor (GR) is a transcription factor that regulates gene expression in a ligand-dependent fashion. This modular protein is one of the major pharmacological targets due to its involvement in both cause and treatment of many human diseases. Intense efforts have been made to get information about the molecular basis of GR activity. Methodology/Principal Findings: Here, the behavior of four GR-ligand complexes with different glucocorticoid and antiglucocorticoid properties were evaluated. The ability of GR-ligand complexes to oligomerize in vivo was analyzed by performing the novel Number and Brightness assay. Results showed that most of GR molecules form homodimers inside the nucleus upon ligand binding. Additionally, in vitro GR-DNA binding analyses suggest that ligand structure modulates GRDNA interaction dynamics rather than the receptor's ability to bind DNA. On the other hand, by coimmunoprecipitation studies we evaluated the in vivo interaction between the transcriptional intermediary factor 2 (TIF2) coactivator and different GR-ligand complexes. No correlation was found between GR intranuclear distribution, cofactor recruitment and the homodimerization process. Finally, Molecular determinants that support the observed experimental GR LBD-ligand/TIF2 interaction were found by Molecular Dynamics simulation. Conclusions/Significance: The data presented here sustain the idea that in vivo GR homodimerization inside the nucleus can be achieved in a DNA-independent fashion, without ruling out a dependent pathway as well. Moreover, since at least one GR-ligand complex is able to induce homodimer formation while preventing TIF2 coactivator interaction, results suggest that these two events might be independent from each other. Finally, 21-hydroxy-6,19-epoxyprogesterone arises as a selective glucocorticoid with potential pharmacological interest. Taking into account that GR homodimerization and cofactor recruitment are considered essential steps in the receptor activation pathway, results presented here contribute to understand how specific ligands influence GR behavior. © 2010 Presman et al.
Fil:Presman, D.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Alvarez, L.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Levi, V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Martí, M.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Veleiro, A.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Burton, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Pecci, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fuente
PLoS ONE 2010;5(10)
Materia
21 hemisuccinate 6,19 epoxyprogesterone
21 hydroxy 6,19 epoxyprogesterone
dexamethasone
glucocorticoid receptor
glucocorticoid receptor antagonist
mifepristone
nuclear receptor coactivator 2
unclassified drug
DNA
glucocorticoid receptor
steroid
article
binding affinity
cell assay
cell culture
cell strain BHK
cell strain COS7
cell strain L 929
cell type
computer model
conformational transition
controlled study
dissociation
DNA drug complex
drug mechanism
drug receptor binding
genetic transfection
in vitro study
in vivo study
incubation time
ligand binding
modulation
molecular dynamics
oligomerization
protein DNA binding
protein DNA interaction
protein function
steroid binding
structure activity relation
animal
cell line
cell strain COS1
Cercopithecus
chemical structure
dimerization
gel mobility shift assay
immunoprecipitation
metabolism
protein binding
Animals
Cell Line
Cercopithecus aethiops
COS Cells
Dimerization
DNA
Electrophoretic Mobility Shift Assay
Immunoprecipitation
Models, Molecular
Molecular Dynamics Simulation
Protein Binding
Receptors, Glucocorticoid
Steroids
Nivel de accesibilidad
Acceso abierto
Licencia
http://creativecommons.org/licenses/by/2.5/ar
Repositorio
Biblioteca Digital (UBA-FCEN)
Institución
Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
OAI Identificador
snrd:HASH013de1e04fe1c7966aedfbab