RRC ID 28454
Author Niwa H, Masui S, Chambers I, Smith AG, Miyazaki J.
Title Phenotypic complementation establishes requirements for specific POU domain and generic transactivation function of Oct-3/4 in embryonic stem cells.
Journal Mol. Cell. Biol.
Abstract Transcription factors of the POU family govern cell fate through combinatorial interactions with coactivators and corepressors. The POU factor Oct-3/4 can define differentiation, dedifferentation, or self-renewal of pluripotent embryonic stem (ES) cells in a sensitive, dose-dependent manner (H. Niwa, J.-I. Miyazali, and A. G. Smith, Nat. Genet. 24:372-376, 2000). Here we have developed a complementation assay based on the ability of Oct-3/4 transgenes to rescue self-renewal in conditionally null ES cells and used this to define which domains of Oct-3/4 are required to sustain the undifferentiated stem cell phenotype. Surprisingly, we found that molecules lacking either the N-terminal or C-terminal transactivation domain, though not both, can effectively replace full-length Oct-3/4. Furthermore, a fusion of the heterologous transactivation domain of Oct-2 to the Oct-3/4 POU domain can also sustain self-renewal. Thus, the unique function of Oct-3/4 in ES cell propagation resides in combination of the specific POU domain with a generic proline-rich transactivation domain. Interestingly, however, Oct-3/4 target gene expression elicited by the N- and C-terminal transactivation domains is not identical, indicating that at least one class of genes activated by Oct-3/4 is not required for ES cell propagation.
Volume 22(5)
Pages 1526-36
Published 2002-3
PMID 11839818
PMC PMC134688
MeSH Animals Cells, Cultured DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism* Embryo, Mammalian / cytology* Genetic Complementation Test Humans Mice Mutation Octamer Transcription Factor-3 Protein Binding Protein Structure, Tertiary Sequence Deletion Stem Cells / physiology* Transcription Factors / genetics Transcription Factors / metabolism* Transcriptional Activation*
IF 3.813
Times Cited 196
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY
Resource
DNA material ptk-luc (RDB07031) pFGF4tk-luc (RDB07032) p6W5'tk-luc (RDB07033) phUTF1tk-luc (RDB07034) pPyCAG-BstXI-IP (RDB07099) pPyCAG-cHA-IP (RDB07100) pBRPyCAG-cHA-IP (RDB07101) pPyCAG-Oct3wt-IRESpuropA (RDB09609) ptk-luc Fgf4enh3' (RDB09711) p052e/p-luc (RDB09725) pRex1p-luc (RDB09723) pmPGKp-luc (RDB09790).