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The mechanism of decrease in nucleolar RNA synthesis by protein synthesis inhibition.

Y Mishima, T Matsui, M Muramatsu

    Journal of Biochemistry
    |March 1, 1979
    PubMed
    Summary
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    Inhibiting protein synthesis rapidly reduces nucleolar RNA synthesis by affecting RNA polymerase IB. A short-lived protein is crucial for maintaining ribosomal RNA transcription and RNA polymerase IB reinitiation.

    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Biochemistry

    Background:

    • Nucleolar RNA synthesis is vital for ribosome biogenesis.
    • Protein synthesis is closely linked to nucleolar function.
    • RNA polymerase I is responsible for ribosomal RNA transcription.

    Purpose of the Study:

    • To investigate the effect of protein synthesis inhibition on nucleolar RNA synthesis.
    • To identify the specific RNA polymerase I forms involved.
    • To elucidate the mechanism of RNA polymerase I regulation in nucleoli.

    Main Methods:

    • Inhibition of protein synthesis using cycloheximide and puromycin.
    • Measurement of nucleolar RNA synthesis.
    • Assay of endogenous RNA polymerase activity in isolated nuclei and nucleoli.

    Related Experiment Videos

  • Fractionation of RNA polymerase I forms using phosphocellulose chromatography.

    • Main Results:

    • Protein synthesis inhibition decreased nucleolar RNA synthesis by ~70% within 1 hour.
    • Only the RNA polymerase IB form decreased in nucleoli, while activity in whole nuclei remained unchanged.
    • Inhibition led to a shift of RNA polymerase IB from bound to free forms in isolated nuclei.
    • Nucleolar RNA polymerase IB activity decreased but recovered rapidly upon resuming protein synthesis.

    Conclusions:

    • A short-lived protein is essential for maintaining ribosomal RNA transcription.
    • This protein facilitates reinitiation of transcription by RNA polymerase IB in the nucleolus.
    • RNA polymerase IB is regulated by protein synthesis, likely through changes in its nucleolar binding or localization.