A protein kinase inhibitor gene reduces both basal and multihormone-stimulated prolactin gene transcription

Richard Day, J. A. Walder, R. A. Maurer

Research output: Contribution to journalArticle

157 Citations (Scopus)

Abstract

The possible role of the catalytic subunit of the cAMP-dependent protein kinase in mediating the regulation of prolactin gene transcription has been investigated through the use of a synthetic gene encoding the heat-stable inhibitor of the cAMP-dependent protein kinase. To assess the effects of protein kinase inhibitor expression on cAMP induction of prolactin gene transcription, a marker gene containing the rat prolactin promoter and adjacent 5'-flanking sequences linked to the bacterial chloramphenicol acetyltransferase gene was cotransfected with a protein kinase inhibitor-expression vector. The results demonstrate that the protein kinase inhibitor-expression vector reduced both basal and cAMP-stimulated expression of the cotransfected prolactin-chloramphenicol acetyltransferase gene. A mutant protein kinase inhibitor-expression vector, coding for an inactive inhibitor protein, did not inhibit basal or cAMP-stimulated prolactin gene transcription. Furthermore, the protein kinase inhibitor-expression vecotr did not inhibit zinc induction of the metallothionein promoter. Analysis of protein kinase activity in transfected cells demonstrated that the protein kinase inhibitor expression vector reduced cAMP-dependent protein kinase activity but did not reduce protein kinase C activity. Nuclease protection experiments confirmed that the effects of the inhibitor vector involved changes in correctly initiated transcripts produced from the prolactin promoter. Surprisingly, the protein kinase inhibitor-expression vector reduced the effects of several different agents including epidermal growth factor, thyrotropin-releasing hormone, phorbol esters, and estrogen on prolactin gene expression to the same extent as it altered cAMP effects.

Original languageEnglish (US)
Pages (from-to)431-436
Number of pages6
JournalJournal of Biological Chemistry
Volume264
Issue number1
StatePublished - 1989
Externally publishedYes

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Transcription
Protein Kinase Inhibitors
Prolactin
Genes
Cyclic AMP-Dependent Protein Kinases
Chloramphenicol O-Acetyltransferase
Synthetic Genes
Gene encoding
Thyrotropin-Releasing Hormone
Metallothionein
5' Flanking Region
Phorbol Esters
Mutant Proteins
Epidermal Growth Factor
Gene expression
Protein Kinases
Protein Kinase C
Zinc
Rats
Catalytic Domain

ASJC Scopus subject areas

  • Biochemistry

Cite this

A protein kinase inhibitor gene reduces both basal and multihormone-stimulated prolactin gene transcription. / Day, Richard; Walder, J. A.; Maurer, R. A.

In: Journal of Biological Chemistry, Vol. 264, No. 1, 1989, p. 431-436.

Research output: Contribution to journalArticle

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