Characterization of a nontissue-specific, 3′,5′-cyclic adenosine monophosphate-responsive element in the proximal region of the rat prolactin gene

Jie Liang, Kyoon E. Kim, William E. Schoderbek, Richard Maurer

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

The nature of a DNA element located in the -100 to -85 region of the rat PRL gene has been characterized. Previous studies demonstrated that this region may contribute to basal and hormonally regulated expression of the PRL gene. As this region contains a sequence with similarity to a consensus cAMP-responsive element (CRE), a possible role for the cAMP response element binding protein (CREB) has been explored. A point mutation which made the PRL CRE-like sequence less like a consensus CRE had little effect on basal or cAMP-stimulated expression of a PRL-luciferase reporter gene. DNase footprint studies demonstrated that the proximal region of the PRL gene does not contain a high affinity CREB binding site. Mobility shift experiments demonstrated that the major GH3 nuclear protein which interacts with the -100 to -85 region of the PRL gene in vitro is not CREB. Transfection of a dominant inhibitor of CREB action had little or no effect on expression of an indicator gene containing the PRL proximal region. Thus, the PRL proximal region does not contain a high affinity CREB binding site, and it is unlikely that CREB plays a major role in expression of the PRL gene. The functional capabilities of the -100 to -85 region of the PRL gene were then tested in a transfection assay. Synthetic multimers of this region were found to be sufficient to permit a transcriptional response to cAMP or TRH in GH3 cells and cAMP in Rat-1 cells. These findings suggest that the proximal region of the rat PRL gene contains a functional, nontissue-specific CRE which likely contributes to the hormonal responsiveness of the PRL gene.

Original languageEnglish (US)
Pages (from-to)885-892
Number of pages8
JournalMolecular Endocrinology
Volume6
Issue number6
StatePublished - Jun 1992
Externally publishedYes

Fingerprint

Cyclic AMP Response Element-Binding Protein
Prolactin
Cyclic AMP
Genes
Gene Expression
Protein Binding
Transfection
Binding Sites
Deoxyribonucleases
Nuclear Proteins
Luciferases
Reporter Genes
Point Mutation
DNA

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Characterization of a nontissue-specific, 3′,5′-cyclic adenosine monophosphate-responsive element in the proximal region of the rat prolactin gene. / Liang, Jie; Kim, Kyoon E.; Schoderbek, William E.; Maurer, Richard.

In: Molecular Endocrinology, Vol. 6, No. 6, 06.1992, p. 885-892.

Research output: Contribution to journalArticle

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