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Related Experiment Videos

Inactive mRNA-protein complexes from mouse sarcoma-180 ascites cells.

T Geoghegan, S Cereghini, G Brawerman

    Proceedings of the National Academy of Sciences of the United States of America
    |November 1, 1979
    PubMed
    Summary

    Mouse sarcoma cells have inactive mRNA in ribonucleoprotein particles, limiting protein synthesis. Cycloheximide treatment partially activates mRNA, but some remains refractory, suggesting regulation by protein complexes.

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    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Biochemistry

    Background:

    • Mouse sarcoma ascites cells exhibit suboptimal protein synthesis capacity.
    • A significant fraction of cellular ribosomes exist as inactive monomers.
    • Substantial cellular messenger RNA (mRNA) is found in 20-70S ribonucleoprotein particles (RNPs).

    Purpose of the Study:

    • To investigate the state and translational activity of mRNA in mouse sarcoma ascites cells.
    • To understand the regulation of protein synthesis and mRNA utilization.
    • To identify factors influencing mRNA availability for translation.

    Main Methods:

    • Sedimentation analysis of ribonucleoprotein particles.
    • Poly(A) content measurements.
    • In vitro translation assays using cell-free systems (reticulocyte lysate).

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  • Treatment with cycloheximide and starvation to modulate cellular conditions.

    • Main Results:

    • RNA from 20-70S RNPs directed the synthesis of less heterogeneous polypeptides compared to polysomal RNA.
    • Cycloheximide treatment promoted ribosomal monomers and some untranslated mRNA into polysomes, but some mRNA remained inactive.
    • RNPs from starved cells were translationally active in vitro, yielding polypeptides similar to polysomal RNA.
    • RNPs from cycloheximide-treated cells were inactive but yielded active RNA after deproteinization.

    Conclusions:

    • Some mRNA species in mouse sarcoma ascites cells are stored in an inactive state within 20-70S RNPs.
    • This inactivity may be mediated by protein components of the nucleoprotein complex.
    • Specific mRNA populations are differentially regulated, with some being refractory to activation treatments.