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Nuclear ribonucleoprotein complexes containing U1 and U2 RNA.

N B Raj, T S Ro-Choi, H Busch

    Biochemistry
    |October 7, 1975
    PubMed
    Summary
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    Researchers isolated nuclear ribonucleoprotein (RNP) complexes containing U1 and U2 RNA from Novikoff hepatoma cells. These complexes contain 10 proteins, with two identified as phosphorylated in vivo.

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Background:

    • Nuclear ribonucleoprotein (RNP) complexes are crucial for gene regulation and RNA processing.
    • Understanding the composition of these complexes provides insights into nuclear functions.

    Purpose of the Study:

    • To isolate and characterize nuclear ribonucleoprotein (RNP) complexes containing U1 and U2 RNA from Novikoff hepatoma cells.
    • To determine the protein composition and phosphorylation status of these RNP complexes.

    Main Methods:

    • Extraction of nuclear RNP complexes using Tris-HCl buffer after washing with NaCl and EDTA.
    • Purification via Sepharose 6B chromatography and sucrose density gradient centrifugation.
    • RNA identification using polyacrylamide gel electrophoresis and T1 RNase fingerprinting.

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  • Protein analysis using two-dimensional polyacrylamide gel electrophoresis.

    • Main Results:

    • Successfully isolated RNP complexes containing U1 and U2 RNA.
    • Confirmed RNA identity through established biochemical methods.
    • Determined a buoyant density of 1.47 g/ml in CsCl gradients for the nuclear RNP complexes.
    • Identified 10 polypeptide spots in the RNP complexes, with two found to be phosphorylated in vivo.

    Conclusions:

    • The study successfully isolated and characterized nuclear RNP complexes containing U1 and U2 RNA.
    • The identified protein components and their phosphorylation provide a basis for further investigation into RNP complex function.
    • These findings contribute to the understanding of nuclear RNP composition and potential roles in cellular processes.