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Related Experiment Videos

F-actin aggregates in transformed cells.

W W Carley, L S Barak, W W Webb

    The Journal of Cell Biology
    |September 1, 1981
    PubMed
    Summary

    Transformed cells exhibit unique actin patches near the cell surface. These actin aggregates appear across various cancer cell types, suggesting a common role in cell transformation and morphology changes.

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    Area of Science:

    • Cell Biology
    • Molecular Biology
    • Cancer Research

    Background:

    • Polymerized actin (F-actin) forms distinct aggregates, or patches, within cultured cells that have undergone transformation.
    • These F-actin patches are consistently observed near the ventral surface of cells transformed by diverse agents, including RNA/DNA tumor viruses, chemical mutagens, and spontaneous transformation.

    Purpose of the Study:

    • To investigate the ubiquity and morphological significance of F-actin aggregates in transformed cells.
    • To determine the temporal relationship between viral oncogene expression and the formation of actin patches.
    • To explore potential mechanisms, including kinase activity, involved in F-actin aggregate formation.

    Main Methods:

    • Utilized nitrobenzoxadiazole-phallacidin, an F-actin-specific fluorescent probe, to visualize and label F-actin aggregates.
    • Examined eight different types of transformed cell lines.
    • Studied actin patch development in normal rat kidney (NRK) cells infected with a temperature-sensitive Rous sarcoma virus mutant (LA23-NRK) following temperature shifts.

    Main Results:

    • F-actin aggregates were identified in all eight transformed cell types examined, indicating their widespread presence.
    • In LA23-NRK cells, actin patches formed rapidly (within 30 minutes) after shifting to the permissive temperature, coinciding with viral src gene expression.
    • Actin aggregates were also observed in cells transformed by non-retroviral agents, suggesting broader involvement beyond viral oncogene activity.

    Conclusions:

    • The ubiquitous presence of F-actin aggregates in transformed cells suggests a common role in mediating or reflecting transformation-associated changes in cell morphology.
    • The rapid appearance of actin patches upon activation of viral src gene expression implicates pp60src kinase activity in cytoskeletal destabilization.
    • The formation of these actin aggregates in non-retrovirally transformed cells indicates that alternative mechanisms, potentially involving endogenous kinases, contribute to this common cellular trait.

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