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Nucleic Acids Research, 1988, Vol. 16, No. 2 487-500
© 1988

Articles

Differential requirements for cellular enhancers in stem and differentiated cells

Thomas F. Vogt, Reid S. Compton1, Richard W. Scott2 and Shirley M. Tilghman1

Institute for Cancer Research Philadelphia, PA 19111 1Department of Molecular Biology, Princeton University Princeton, NJ 08544 2E.I.du Pont de Nemours and Company, Inc., Central Research and Development Department Experimental Station, E328/140, Wilmington, DE 19898, USA

Received October 20, 1987. Revised December 14, 1987. Accepted December 14, 1987.

Multiple cis-acting regulatory elements consisting of three cellular enhancers and a proximal promoter element have been identified in the region upstream of the mouse {alpha}-fetoprotein (AFP) gene. We examined the role of these sequences during differentiation by the introduction of modified AFP genes into cells at different stages of commitment to its expression. Modified AFP genes introduced stably into F9 embryonal carcinoma stem cells by DNA transfection were silent until activated by treatment with retinoic acid to form visceral endoderm. Their activation required the presence of both the enhancer and proximal promoter domains. The introduced genes activated simultaneously with the endogenous AFP genes, but reached maximal levels of expression more rapidly, suggesting a greater initial accessibility to transcription factors. In contrast, when aodified AFP genes were stably introduced into HepG2 cells, a human hepatoraa cell line that constitutively expresses the AFP gene, the proximal promoter sequences were sufficient to direct a low level of expression. The absolute requirement for the AFP enhancers in F9 cells but not in HepG2 cells supports a model by which there is an obligate requirement for enhancers during differentiation in addition to their role in enhancing gene expression after differentiation.


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