Combination of Runx2 and Cbfß upregulates Amelotin gene expression in ameloblasts by directly interacting with cisenhancers during amelogenesis.
Mol Med Rep
; 17(4): 6068-6076, 2018 04.
Article
en En
| MEDLINE
| ID: mdl-29436627
ABSTRACT
Amelotin (Amtn) is a recently identified enamel protein secreted by ameloblasts at late stage of enamel development. Runtrelated transcription factor 2 (Runx2) in combination with the coactivator corebinding factor ß (Cbfß) regulates the early stages of tooth development. The aim of the present study was to investigate the role of Runx2 in the regulation of Amtn gene expression in ameloblasts. Immunohistochemistry was performed and the results revealed that Runx2 protein was predominantly expressed in the nuclei of ameloblasts during the transition stage and the maturation stage of enamel development, whereas Cbfß was expressed in ameloblasts from the secretory stage to the maturation stage. Reverse transcriptionquantitative polymerase chain reaction results demonstrated that Runx2 knockdown decreased Amtn expression in ameloblastlineage cells and coexpression of Runx2 and Cbfß in ameloblast lineage cells induced an upregulation in Amtn gene expression. Two putative Runx2binding sites within the Amtn promoter were identified using bioinformatics analysis. Results of an electrophoretic mobility shift assay and chromatin immunoprecipitation indicated that Runx2/Cbfß bound to specific DNA sequences. Sitedirected mutagenesis of the Runx2 binding sites within the Amtn promoter resulted in decreased basal promoter activity and did not affect the overexpressed Runx2/Cbfß. The results of the present study suggest that Runx2 upregulates Amtn gene expression via binding directly to Runx2 sites within the Amtn promoter during amelogenesis.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Regulación de la Expresión Génica
/
Elementos de Facilitación Genéticos
/
Factor de Unión a CCAAT
/
Proteínas del Esmalte Dental
/
Subunidad alfa 1 del Factor de Unión al Sitio Principal
/
Ameloblastos
/
Amelogénesis
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
Mol Med Rep
Año:
2018
Tipo del documento:
Article