RRC ID 31699
Author Okumura-Nakanishi S, Saito M, Niwa H, Ishikawa F.
Title Oct-3/4 and Sox2 regulate Oct-3/4 gene in embryonic stem cells.
Journal J. Biol. Chem.
Abstract Oct-3/4 is a key transcriptional factor whose expression level governs the fate of primitive inner cell mass and embryonic stem (ES) cells. Previously, an upstream 3.3-kb distal enhancer (DE) fragment was identified to be responsible for the specific expression of mouse Oct-3/4 in the inner cell mass and ES cells. However, little is known about the cis-elements and trans-factors required for DE activity. In this study, we identified a novel cis-element, called Site 2B here, located approximately 30 bp downstream from Site 2A, which was previously revealed in DE by an in vivo chemical modification experiment. Using the luciferase reporter assay, we demonstrated that both Site 2A and Site 2B are necessary and sufficient for activating DE in the contexts of both the native Oct-3/4 promoter and the heterologous thymidine kinase minimal promoter. In an electrophoretic mobility shift assay we showed that Site 2B specifically binds to Oct-3/4 and Sox2 when ES-derived cell extracts were used, whereas Site 2A binds to a factor(s) present in both ES and NIH 3T3 cells. Furthermore, we showed that the physiological level of Oct-3/4 in ES cells is required for Site 2B-mediated DE activity using the inducible knock-out system of Oct-3/4 in ES cells. These results indicate that Oct-3/4 is a member of the gene family regulated by Oct-3/4 and Sox2, as reported before for the FGF-4, UTF1, Sox2, and Fbx15 genes. Thus, Oct-3/4 and Sox2 comprise a regulatory complex that controls the expression of genes important for the maintenance of the primitive state, including themselves. This autoregulatory circuit of the Sox2.Oct-3/4 complex may contribute to maintaining robustly the precise expression level of Oct-3/4 in primitive cells.
Volume 280(7)
Pages 5307-17
Published 2005-2-18
DOI 10.1074/jbc.M410015200
PII M410015200
PMID 15557334
MeSH Animals Base Sequence Cell Differentiation / drug effects Cell Line DNA-Binding Proteins / genetics* DNA-Binding Proteins / metabolism* Down-Regulation / drug effects Embryo, Mammalian / cytology* Enhancer Elements, Genetic / genetics Gene Expression Regulation, Developmental* / drug effects Humans Mice Molecular Sequence Data Mutation / genetics NIH 3T3 Cells Octamer Transcription Factor-3 Promoter Regions, Genetic / genetics Response Elements / genetics SOXB1 Transcription Factors Stem Cells / cytology Stem Cells / metabolism* Trans-Activators / metabolism* Transcription Factors / genetics* Transcription Factors / metabolism* Tretinoin / pharmacology
IF 4.011
Times Cited 226
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material Oct3pluc BstXI-NcoI (RDB10752) Oct3pluc PstI-NcoI (RDB10753) Oct3pluc BamHI-NcoI (RDB10754) pGL3b Hind/NcoI (-1938/-1) (RDB10986).