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Related Experiment Videos

Phosphoinositide kinase activity and transformation.

M Whitman, D Kaplan, L Cantley

    Federation Proceedings
    |October 1, 1986
    PubMed
    Summary

    Polyoma virus middle T antigen activates phosphatidylinositol (PI) kinase activity, crucial for cell transformation. This PI kinase modulation, linked to pp60 c-src, highlights a key pathway in viral oncogenesis.

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    Area of Science:

    • Virology
    • Molecular Biology
    • Cellular Biology

    Background:

    • The phosphatidylinositol (PI) turnover pathway is essential for cellular signaling.
    • DNA tumor viruses, like polyoma, utilize host cell machinery for replication and transformation.
    • The role of PI pathway modulation in viral transformation is not fully understood.

    Purpose of the Study:

    • To investigate if the PI turnover pathway is a critical target for polyoma virus transforming gene products.
    • To determine the association between PI kinase activity and the transforming ability of polyoma virus.
    • To elucidate the mechanism by which polyoma virus middle T antigen influences PI metabolism.

    Main Methods:

    • Utilized polyoma virus as a model system for studying viral transformation.
    • Analyzed PI turnover by measuring polyphosphoinositides and polyphosphoinositols in infected cells.
    • Associated PI kinase activity with middle T antigen using immunoprecipitation assays.
    • Investigated the interaction between middle T antigen and pp60 c-src tyrosine kinase.
    • Examined the effect of tyrosine phosphatase inhibitor (ammonium orthovanadate) on PI kinase activity.

    Main Results:

    • Polyoma-infected cells exhibited elevated levels of polyphosphoinositides and polyphosphoinositols.
    • PI kinase activity was found to be associated with polyoma virus middle T antigen.
    • A strong correlation was observed between middle T-associated PI kinase activity and the virus's transforming ability.
    • Middle T antigen associates with pp60 c-src at the plasma membrane, activating its tyrosine kinase function.
    • Ammonium orthovanadate treatment enhanced the PI kinase activity associated with the middle T/pp60 c-src complex.

    Conclusions:

    • Polyoma virus middle T antigen directly influences the PI turnover pathway.
    • The middle T/pp60 c-src complex plays a significant role in activating PI kinase activity.
    • Tyrosine phosphorylation mediated by the middle T/pp60 c-src complex is proposed to modulate PI turnover in vivo, contributing to viral transformation.

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