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PMID: 11839818 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Phenotypic complementation establishes requirements for specific POU domain and generic transactivation function of Oct-3/4 in embryonic stem cells.

Molecular and cellular biology ·Vol. 22 ·No. 5 ·2002-03-00 ·Pages 1526-36

Niwa H, Masui S, Chambers I, Smith AG, Miyazaki J

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.

MeSH Terms
Animals Cells, Cultured DNA-Binding Proteins/genetics,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,metabolism Transcriptional Activation
Chemicals
DNA-Binding Proteins Octamer Transcription Factor-3 POU5F1 protein, human Pou5f1 protein, mouse Transcription Factors
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Niwa Hitoshi
Stem Cell Regulation Research, Area of Molecular Therapeutics, Course of Advanced Medicine, Osaka University Graduate School of Medicine, Suita C, Osaka 565-0871, Japan. niwa@rtc.riken.go.jp
Masui Shinji
Chambers Ian
Smith Austin G
Miyazaki Jun-ichi
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.22.5.1526-1536.2002
Email
niwa@rtc.riken.go.jp
Published
2002-03-00
Pages
1526-36
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC134688
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.22.5.1526-1536.2002 PMC Full Text View journal details
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