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Molecular Pharmacology

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Research ArticleArticle

Role for Early Growth Response-1 Protein in α1-Adrenergic Stimulation of Fibroblast Growth Factor-2 Promoter Activity in Cardiac Myocytes

Yan Jin, Farah Sheikh, Karen A. Detillieux and Peter A. Cattini
Molecular Pharmacology May 2000, 57 (5) 984-990;
Yan Jin
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Farah Sheikh
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Karen A. Detillieux
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Peter A. Cattini
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Abstract

Fibroblast growth factor-2 (FGF-2), a mitogenic, angiogenic, and cardioprotective agent, is released from the postnatal heart by a mechanism of transient remodelling of the sarcolemma during contraction. Both release of FGF-2 and its synthesis can be increased with adrenergic stimulation. We reported previously that FGF-2 synthesis can be regulated at the transcriptional level by α-adrenergic stimulation of cultured neonatal rat cardiac myocytes as well as in the adult mouse heart. Examination of the proximal promoter region of both human and rat FGF-2 gene sequences revealed binding sites for the early growth response-1 (Egr-1) protein. Using gel mobility shift assays, we observed a transient increase in a complex between nuclear extracts from neonatal rat cardiac myocytes treated with inducers of Egr-1, including the α-adrenergic agonist phenylephrine, angiotensin II, and phorbol ester, and a consensus Egr-1 DNA element. A similar complex was seen with the FGF-2 promoter region −7/+42 as the DNA probe, but not when the Egr-1 element at nucleotides +3/+31 was disrupted. Participation of Egr-1 protein in the complex was confirmed by competition with Egr-1 DNA elements and antibodies. With deletion analysis and transfection of neonatal rat cardiac myocytes, the α-adrenergic response was localized to nucleotides −110/+42 of the FGF-2 gene in the context of a hybrid FGF-2/luciferase reporter gene, −110FGFp.luc. Overexpression of Egr-1 increased −110FGFp.luc gene expression, whereas mutation of its Egr-1 element at nucleotides +3/+31 abolished α-adrenergic responsiveness. These data indicate that Egr-1 is involved in the α-adrenergic stimulation of the FGF-2 promoter region in neonatal cardiac myocytes.

Footnotes

  • Send reprint requests to: Peter A. Cattini, Department of Physiology, University of Manitoba, 730 William Ave., Winnipeg, Manitoba, Canada R3E 3J7. E-mail: Peter_Cattini{at}UManitoba.CA

  • This study was supported by a grant from the Medical Research Council of Canada (to M.R.C.). F.S. was the recipient of an Medical Research Council Studentship, K.A.D. was the recipient of a Heart and Stroke Foundation Studentship, and P.A.C. was the recipient of a Medical Research Council Scientist award.

  • Abbreviations:
    NE
    norepinephrine
    FGF-2
    fibroblast growth factor-2
    Egr-1
    early growth response-1
    FBS
    fetal bovine serum
    CMV
    cytomegalovirus
    SV40
    simian virus 40
    EMSA
    electrophoretic gel mobility shift assay
    PMA
    phorbol-12-myristate-13-acetate
    ATII
    angiotensin II
    WT
    wild type
    MUT
    mutation
    PE
    phenylephrine
    DMEM
    Dulbecco's modified Eagle's medium
    Praz
    prazosin
    • Received November 29, 1999.
    • Accepted February 2, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 57 (5)
Molecular Pharmacology
Vol. 57, Issue 5
1 May 2000
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Research ArticleArticle

Role for Early Growth Response-1 Protein in α1-Adrenergic Stimulation of Fibroblast Growth Factor-2 Promoter Activity in Cardiac Myocytes

Yan Jin, Farah Sheikh, Karen A. Detillieux and Peter A. Cattini
Molecular Pharmacology May 1, 2000, 57 (5) 984-990;

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Research ArticleArticle

Role for Early Growth Response-1 Protein in α1-Adrenergic Stimulation of Fibroblast Growth Factor-2 Promoter Activity in Cardiac Myocytes

Yan Jin, Farah Sheikh, Karen A. Detillieux and Peter A. Cattini
Molecular Pharmacology May 1, 2000, 57 (5) 984-990;
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