The progesterone receptor stimulates cell-free transcription by enhancing the formation of a stable preinitiation complex

Ludger Klein-Hitpass, Sophia Y. Tsai, Nancy L. Weigel, George F. Allan, Deborah Riley, Ronald Rodriguez, William T. Schrader, Ming ler Tsai, Bert W. O'Malley

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Abstract

Highly purified chicken progesterone receptor (cPR) is shown to stimulate RNA synthesis directly in an in vitro transcription assay. Stimulation of transcription by cPR requires the presence of progesterone response elements (PREs) in the template and can be specifically inhibited by addition of competitor oligonucleotides containing PREs. Binding of receptor to two PREs is cooperative and leads to a synergistic (27-fold) stimulation of transcription. A purified fusion protein containing the DNA binding domain of cPR linked to yeast ubiquitin was produced in E. coli and also functions in the transcription assay. Using this in vitro transcription system, we demonstrate that hormone-free cPR activated by salt treatment induces transcription of a test gene in a hormone-independent manner. Finally, we present evidence that the progesterone receptor acts by facilitating the formation of a stable preinitiation complex at the target gene promoter and thus augments the initiation of transcription by RNA polymerase II.

Original languageEnglish (US)
Pages (from-to)247-257
Number of pages11
JournalCell
Volume60
Issue number2
DOIs
StatePublished - Jan 26 1990
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Klein-Hitpass, L., Tsai, S. Y., Weigel, N. L., Allan, G. F., Riley, D., Rodriguez, R., Schrader, W. T., Tsai, M. L., & O'Malley, B. W. (1990). The progesterone receptor stimulates cell-free transcription by enhancing the formation of a stable preinitiation complex. Cell, 60(2), 247-257. https://doi.org/10.1016/0092-8674(90)90740-6