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Related Experiment Videos

Nucleoprotein complexes containing replicating Simian virus 40 DNA: comparison with polyoma nucleoprotein complexes.

M R Hall, W Meinke, D A Goldstein

    Journal of Virology
    |October 1, 1973
    PubMed
    Summary

    Researchers isolated replicating simian virus 40 (SV40) DNA-protein complexes from infected cells. These complexes, unlike those with supercoiled DNA, showed faster sedimentation, indicating distinct structural properties of replicating SV40 nucleoproteins.

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    Area of Science:

    • Molecular Biology
    • Virology
    • Biochemistry

    Background:

    • Nucleoprotein complexes are crucial for viral DNA replication.
    • Understanding the structure of replicating viral DNA within host cells is essential.

    Purpose of the Study:

    • To isolate and characterize short-lived nucleoprotein complexes containing replicating simian virus 40 (SV40) DNA.
    • To compare the properties of replicating SV40 nucleoprotein complexes with those containing supercoiled SV40 and polyoma DNA.

    Main Methods:

    • Isolation of nucleoprotein complexes from infected monkey cells using cell lysis and Triton X-100 extraction.
    • Sucrose gradient sedimentation to determine the sedimentation rates of various SV40 and polyoma DNA complexes.
    • Buoyant density measurements to assess protein-to-DNA ratios.

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    Main Results:

    • Replicating SV40 (r-SV40) complexes were isolated and shown to be partially nuclear-bound.
    • r-SV40 complexes exhibited faster sedimentation rates (56-75S) compared to SV40 complexes (61S) and polyoma complexes (56S).
    • Sedimentation and density analyses indicated a similar protein-to-DNA ratio (1-2/1) in SV40 nucleoprotein complexes as previously reported for polyoma nucleoproteins.

    Conclusions:

    • Replicating SV40 DNA exists in distinct nucleoprotein complexes with unique sedimentation properties.
    • The protein association with SV40 DNA is consistent across different forms, suggesting conserved packaging mechanisms.