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Related Experiment Videos

The ribonucleoprotein substrate for a ribosomal RNA-processing nuclease.

D A Stahl, B Pace, T Marsh

    The Journal of Biological Chemistry
    |September 25, 1984
    PubMed
    Summary

    Bacillus subtilis RNase M5, crucial for 5S rRNA maturation, involves proteins alpha and beta. Ribosomal protein BL16 (beta component) binds 5S rRNA, with electrostatic interactions dominating the binding energy.

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    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Ribosome Biogenesis

    Background:

    • RNase M5 is essential for the endonucleolytic maturation of 5S ribosomal RNA (rRNA) in Bacillus subtilis.
    • This activity requires two protein components, designated alpha and beta.
    • The beta component has been identified as ribosomal protein BL16.

    Purpose of the Study:

    • To characterize the role of Bacillus subtilis ribosomal protein BL16 in 5S rRNA maturation.
    • To investigate the binding interactions between BL16 and 5S rRNA, including precursor forms.
    • To determine the contribution of electrostatic forces to the BL16-5S rRNA complex formation.

    Main Methods:

    • Purification of the beta component (BL16) to homogeneity.
    • Functional complementation assays using Bacillus subtilis alpha protein and Escherichia coli ribosomal protein EL18.

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  • Filter binding assays to evaluate the association of BL16 with various RNA molecules.
  • Ionic strength-dependent analysis to assess electrostatic contributions to binding.

    • Main Results:

    • Bacillus subtilis ribosomal protein BL16 was purified and confirmed as the beta component of RNase M5.
    • BL16 functionally complements the alpha component, similar to E. coli EL18.
    • BL16 binds mature 5S rRNA and its native precursor with similar affinity; precursor sequences do not enhance binding.
    • Electrostatic interactions account for approximately 65% of the binding energy between BL16 and 5S rRNA.

    Conclusions:

    • Ribosomal protein BL16 is the functional beta component of Bacillus subtilis RNase M5.
    • The maturation precursor sequences of 5S rRNA do not facilitate BL16 binding.
    • Electrostatic forces play a significant role in the association of ribosomal protein BL16 with 5S rRNA.