Characterization of DNA regions mediating the ability of Ca2+/calmodulin dependent protein kinase II to stimulate prolactin promoter activity

B. E. Nowakowski, Y. Okimura, Richard Maurer

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The ability of Ca2+/calmodulin-dependent protein kinases (CaMKs) to regulate transcription of the rat prolactin (PRL) gene has been examined. We found that KN-62, a potent inhibitor of CaM kinases, blunted the ability of TRH to activate the prolactin promoter. Transfection experiments using expresssion plasmids for constitutively active forms of CaMKI, CaMKII, or CaMKIV show that CaMKII is the most effective activator of prolactin promoter expression. Deletion studies demonstrated that the upstream boundary of sequences necessary to respond to CaMKII is located within the distal enhancer of the prolactin gene. Neither the distal enhancer alone nor the proximal region of the prolactin gene are sufficient to mediate a response to CaMKII. Mutational analysis suggests that several Pit-1 binding sites contribute to CaMKII responsiveness. These findings suggest that CaMKII responsiveness of the prolactin promoter requires multiple factor binding sites in both the distal and proximal regions of the gene.

Original languageEnglish (US)
Pages (from-to)109-116
Number of pages8
JournalMolecular and Cellular Endocrinology
Volume132
Issue number1-2
DOIs
StatePublished - 1997

Fingerprint

Calcium-Calmodulin-Dependent Protein Kinase Type 2
Prolactin
DNA
Genes
KN 62
Binding Sites
Calcium-Calmodulin-Dependent Protein Kinases
Transcription
Transfection
Rats
Plasmids
Phosphotransferases

Keywords

  • Calcium
  • Prolactin
  • Protein kinase
  • Transcription

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

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abstract = "The ability of Ca2+/calmodulin-dependent protein kinases (CaMKs) to regulate transcription of the rat prolactin (PRL) gene has been examined. We found that KN-62, a potent inhibitor of CaM kinases, blunted the ability of TRH to activate the prolactin promoter. Transfection experiments using expresssion plasmids for constitutively active forms of CaMKI, CaMKII, or CaMKIV show that CaMKII is the most effective activator of prolactin promoter expression. Deletion studies demonstrated that the upstream boundary of sequences necessary to respond to CaMKII is located within the distal enhancer of the prolactin gene. Neither the distal enhancer alone nor the proximal region of the prolactin gene are sufficient to mediate a response to CaMKII. Mutational analysis suggests that several Pit-1 binding sites contribute to CaMKII responsiveness. These findings suggest that CaMKII responsiveness of the prolactin promoter requires multiple factor binding sites in both the distal and proximal regions of the gene.",
keywords = "Calcium, Prolactin, Protein kinase, Transcription",
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T1 - Characterization of DNA regions mediating the ability of Ca2+/calmodulin dependent protein kinase II to stimulate prolactin promoter activity

AU - Nowakowski, B. E.

AU - Okimura, Y.

AU - Maurer, Richard

PY - 1997

Y1 - 1997

N2 - The ability of Ca2+/calmodulin-dependent protein kinases (CaMKs) to regulate transcription of the rat prolactin (PRL) gene has been examined. We found that KN-62, a potent inhibitor of CaM kinases, blunted the ability of TRH to activate the prolactin promoter. Transfection experiments using expresssion plasmids for constitutively active forms of CaMKI, CaMKII, or CaMKIV show that CaMKII is the most effective activator of prolactin promoter expression. Deletion studies demonstrated that the upstream boundary of sequences necessary to respond to CaMKII is located within the distal enhancer of the prolactin gene. Neither the distal enhancer alone nor the proximal region of the prolactin gene are sufficient to mediate a response to CaMKII. Mutational analysis suggests that several Pit-1 binding sites contribute to CaMKII responsiveness. These findings suggest that CaMKII responsiveness of the prolactin promoter requires multiple factor binding sites in both the distal and proximal regions of the gene.

AB - The ability of Ca2+/calmodulin-dependent protein kinases (CaMKs) to regulate transcription of the rat prolactin (PRL) gene has been examined. We found that KN-62, a potent inhibitor of CaM kinases, blunted the ability of TRH to activate the prolactin promoter. Transfection experiments using expresssion plasmids for constitutively active forms of CaMKI, CaMKII, or CaMKIV show that CaMKII is the most effective activator of prolactin promoter expression. Deletion studies demonstrated that the upstream boundary of sequences necessary to respond to CaMKII is located within the distal enhancer of the prolactin gene. Neither the distal enhancer alone nor the proximal region of the prolactin gene are sufficient to mediate a response to CaMKII. Mutational analysis suggests that several Pit-1 binding sites contribute to CaMKII responsiveness. These findings suggest that CaMKII responsiveness of the prolactin promoter requires multiple factor binding sites in both the distal and proximal regions of the gene.

KW - Calcium

KW - Prolactin

KW - Protein kinase

KW - Transcription

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