Characterization of a negative retinoic acid response element in the murine Oct4 promoter
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| Published in | Molecular and Cellular Biology Vol. 14; no. 2; pp. 1122 - 1136 |
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| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
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Washington, DC
American Society for Microbiology
01.02.1994
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| ISSN | 0270-7306 1098-5549 |
| DOI | 10.1128/MCB.14.2.1122 |
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Expression of Oct4 in embryonic stem cells is controlled by a distal upstream stem cell-specific enhancer that is deactivated during retinoic acid (RA)-induced differentiation by an indirect mechanism not involving binding of RA receptors. We report that in RA-treated P19 embryonal carcinoma cells the Oct4 promoter is also subject to negative regulation by RA. The minimal Oct4 promoter sequence mediating repression consists of a promoter-proximal sequence containing a GC-rich SP1 consensus-like sequence and several hormone response element half-sites that can be arranged into direct repeats with different spacing. The GC box binds a nuclear factor that is invariably present in undifferentiated and RA-treated differentiated P19 cells. By contrast, the hormone response element-containing sequence binds factors that are induced following RA treatment. Mutational analysis and competition experiments show that the functional entity binding the RA-induced factor is a direct repeat sequence with a spacing of one nucleotide, previously shown to be a binding site for COUP transcription factors (COUP-TFs). Cotransfected orphan receptors COUP-TF1, ARP-1, and EAR-2 were able to repress the activity of Oct4 promoter-driven reporters in P19 EC cells, albeit with different efficiencies. Furthermore, the negative transcriptional effect of COUP-TFs is dominant over the activating effect of the Oct4 embryonic stem cell-specific enhancer. The results show that negative regulation of Oct4 expression during RA-induced differentiation of embryonic stem cells is controlled by two different mechanisms, including deactivation of the embryonic stem cell-specific enhancer and promoter silencing by orphan nuclear hormone receptors. Expression of Oct4 in embryonic stem cells is controlled by a distal upstream stem cell-specific enhancer that is deactivated during retinoic acid (RA)-induced differentiation by an indirect mechanism not involving binding of RA receptors (H. Okazawa, K. Okamoto, F. Ishino, T. Ishino-Kaneko, S. Takeda, Y. Toyoda, M. Muramatsu, and H. Hamada, EMBO J. 10:2997-3005, 1991). Here we report that in RA-treated P19 embryonal carcinoma cells the Oct4 promoter is also subject to negative regulation by RA. The minimal Oct4 promoter sequence mediating repression consists of a promoter-proximal sequence containing a GC-rich SP1 consensus-like sequence and several hormone response element half-sites that can be arranged into direct repeats with different spacing. The GC box binds a nuclear factor that is invariably present in undifferentiated and RA-treated differentiated P19 cells. By contrast, the hormone response element-containing sequence binds factors that are induced following RA treatment. Mutational analysis and competition experiments show that the functional entity binding the RA-induced factor is a direct repeat sequence with a spacing of one nucleotide, previously shown to be a binding site for COUP transcription factors (COUP-TFs). Cotransfected orphan receptors COUP-TF1, ARP-1, and EAR-2 were able to repress the activity of Oct4 promoter-driven reporters in P19 EC cells, albeit with different efficiencies. Furthermore, the negative transcriptional effect of COUP-TFs is dominant over the activating effect of the Oct4 embryonic stem cell-specific enhancer. These results show that negative regulation of Oct4 expression during RA-induced differentiation of embryonic stem cells is controlled by two different mechanisms, including deactivation of the embryonic stem cell-specific enhancer and promoter silencing by orphan nuclear hormone receptors. |
| Author | L Jonk C Pals J Schoorlemmer M van Den Eijnden A van Puijenbroek W Kruijer |
| AuthorAffiliation | Hubrecht Laboratorium, Netherlands Institute for Developmental Biology, Utrecht |
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| Keywords | Ligand binding Vertebrata Mammalia Transcription Mouse Transcription promoter Rodentia Regulatory sequence Transcription factor Cell differentiation Retinoic acid Biological receptor |
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Mendeley... Expression of Oct4 in embryonic stem cells is controlled by a distal upstream stem cell-specific enhancer that is deactivated during retinoic acid (RA)-induced... |
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| SubjectTerms | Animals Base Sequence Binding Sites Biological and medical sciences Cell Differentiation Cell Line Consensus Sequence DNA-Binding Proteins - biosynthesis DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism Embryo, Mammalian Fundamental and applied biological sciences. Psychology Genes, Regulator - drug effects Genomic Library Mice Mice, Inbred CBA Mice, Inbred Strains Molecular and cellular biology Molecular genetics Molecular Sequence Data Octamer Transcription Factor-3 Oligodeoxyribonucleotides Promoter Regions, Genetic - drug effects Repetitive Sequences, Nucleic Acid Restriction Mapping Sequence Homology, Nucleic Acid Spleen - metabolism Stem Cells - metabolism Transcription Factors - genetics Transcription, Genetic Transcription. Transcription factor. Splicing. Rna processing Transfection Tretinoin - pharmacology |
| Title | Characterization of a negative retinoic acid response element in the murine Oct4 promoter |
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