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Related Experiment Videos

A cyclic AMP- and phorbol ester-inducible DNA element.

M Comb, N C Birnberg, A Seasholtz

    Nature
    |September 1, 1986
    PubMed
    Summary
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    Cellular signaling pathways involving cyclic adenosine monophosphate (cAMP) and phorbol esters directly influence proenkephalin gene expression. A specific 37-base pair DNA region is critical for this regulation, acting as a transcriptional enhancer.

    Area of Science:

    • Molecular biology
    • Cell signaling
    • Neuroscience

    Background:

    • Cellular processes are modulated by hormones and neurotransmitters acting via cell-surface receptors and second messengers.
    • Cyclic adenosine monophosphate (cAMP) is a key second messenger regulating adenylate cyclase activity.
    • Phorbol esters activate protein kinase C, involved in the phosphatidylinositol pathway.

    Purpose of the Study:

    • To investigate the regulation of proenkephalin gene expression by second messengers.
    • To identify the DNA sequences responsible for mediating transcriptional regulation by cAMP and phorbol esters.

    Main Methods:

    • Transient expression of a proenkephalin-chloramphenicol acetyl transferase fusion gene in tissue culture cells.
    • Treatment with cAMP analogues, adenylate cyclase activators, and phorbol esters.

    Related Experiment Videos

  • Deletion analysis to map regulatory DNA sequences.

    • Main Results:

    • cAMP analogues and adenylate cyclase activators regulate the fusion gene.
    • Phorbol esters also regulate the fusion gene, particularly with phosphodiesterase inhibitors.
    • A 37-base pair region 5' to the proenkephalin gene's mRNA cap site is essential for regulation by both signaling pathways.
    • This region contains three related 12-bp sequences and functions as a transcriptional enhancer.

    Conclusions:

    • The proenkephalin gene's expression is subject to regulation by intracellular signaling pathways involving cAMP and protein kinase C.
    • A conserved enhancer element within the proenkephalin gene promoter mediates responses to these signaling pathways.
    • This finding elucidates mechanisms of trans-synaptic regulation of gene expression.