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Related Experiment Videos

Cross-linked informofers.

V V Prosvirnin, S Ruzidic, O P Samarina

    Nucleic Acids Research
    |November 24, 1979
    PubMed
    Summary
    This summary is machine-generated.

    Researchers identified informofers, stable protein particles within hnRNP, using cross-linking. These informofers, composed of informatin, can rebind pre-mRNA to reconstitute RNP particles after high salt dissociation.

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    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Cell Biology

    Background:

    • Heterogeneous nuclear ribonucleoprotein (hnRNP) particles are crucial for pre-mRNA processing and transport.
    • The proteinaceous core of these particles, known as informofers, has been challenging to isolate and characterize due to its association with RNA.

    Purpose of the Study:

    • To investigate the proteinaceous core (informofer) of 30S hnRNP particles.
    • To determine if informofers can be isolated and characterized independently of RNA.
    • To assess the ability of isolated informofers to rebind pre-mRNA.

    Main Methods:

    • Cross-linking of hnRNP proteins using bifunctional reagents (dimethyl-suberimidate, dimethyl-3,3'-dithiobispropionimidate).
    • High salt (1-2 M NaCl) treatment to dissociate RNA from proteins.

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  • Analysis of particle stability and composition using sedimentation coefficients.
  • Dissociation of cross-linked informofers using SDS, urea, and beta-mercaptoethanol.
  • Reconstitution of RNP particles by mixing informofers with pre-mRNA.

    • Main Results:

    • Cross-linked informofers, the proteinaceous core of 30S hnRNP, survived high salt treatment that dissociated all RNA.
    • Isolated informofers showed sedimentation coefficients similar to original hnRNP particles and contained no detectable RNA.
    • Informofer dissociation yielded oligomeric polypeptides, identified as informatin.
    • Cross-linked informofers could be reconstituted with pre-mRNA after high salt removal, forming RNP particles.

    Conclusions:

    • Informofers represent a stable, RNA-free protein scaffold within hnRNPs.
    • The protein informatin is the fundamental subunit of informofers.
    • Informofers are capable of binding and organizing pre-mRNA, suggesting a key role in RNA processing and transport.