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Effect of adenosine 3':5'-monophosphate and guanosine 3':5'-monophosphate on RNA release from isolated nuclei

D E Schumm, T E Webb

    The Journal of Biological Chemistry
    |December 10, 1978
    PubMed

    Insights

    Cyclic nucleotides like cAMP and cGMP stimulate messenger RNA release from rat liver nuclei in a cell-free system. This RNA release is dependent on the in vivo levels of these cyclic nucleotides.

    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Biochemistry

    Background:

    • Cyclic nucleotides, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), are crucial intracellular signaling molecules.
    • Regulation of gene expression involves the control of RNA metabolism, including RNA release from the nucleus.

    Purpose of the Study:

    • To investigate the role of cAMP and cGMP in regulating RNA release from isolated rat liver nuclei.
    • To determine if cyclic nucleotides can stimulate the release of specific RNA types, such as messenger RNA (mRNA).

    Main Methods:

    • Utilizing a cell-free system with isolated, prelabeled rat liver nuclei and fortified cytosol.
    • Assessing RNA release upon addition of physiological concentrations of cAMP or cGMP.
    • Characterizing the released RNA using oligo(dT)-cellulose binding and sedimentation analysis.
    • Comparing the response in cell-free systems prepared from different tissues (resting liver, hepatomas, regenerating liver) and correlating with in vivo cyclic nucleotide levels.

    Main Results:

    • Both cAMP and cGMP significantly stimulated the release of RNA from isolated nuclei into the cytosol.
    • The released RNA was predominantly mRNA, confirmed by its binding to oligo(dT)-cellulose and its sedimentation profile.
    • Cyclic nucleotide stimulation of RNA release was observed in systems from resting rat liver, Novikoff hepatoma, and Morris hepatoma 5123D, but not in 18-h regenerating liver.
    • The responsiveness of the cell-free system to added cyclic nucleotides inversely correlated with the endogenous levels of these nucleotides in the source tissue.
    • Neither cAMP nor cGMP affected the release of ribosomal RNA (rRNA).

    Conclusions:

    • Cyclic nucleotides (cAMP and cGMP) play a role in stimulating mRNA release from the nucleus.
    • The regulation of mRNA release by cyclic nucleotides is tissue-specific and influenced by endogenous nucleotide levels.
    • This cell-free system provides a model to study the mechanisms of cyclic nucleotide-mediated RNA release and gene expression regulation.

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