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Micromanipulation Techniques Allowing Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators
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Early changes in the distribution and organization of microfilament proteins during cell transformation

C B Boschek, B M Jockusch, R R Friis

    Cell
    |April 1, 1981
    PubMed

    Insights

    Rous sarcoma virus infection causes early cell surface "flowers" rich in actin and associated proteins, indicating rapid microfilament reorganization during transformation.

    Area of Science:

    • Cell Biology
    • Virology
    • Molecular Biology

    Background:

    • Rous sarcoma virus (RSV) infection induces significant morphological changes in host cells.
    • The viral src gene plays a critical role in mediating these transformation-associated cellular alterations.

    Purpose of the Study:

    • To investigate the early cellular events and protein accumulations during RSV-induced cell transformation.
    • To characterize the composition and origin of early morphological changes, specifically "flowers".

    Main Methods:

    • Utilized temperature-downshift experiments with transformation-defective temperature-sensitive RSV mutants in chick embryo cells.
    • Analyzed cellular morphology and stained for cytoskeletal proteins, including actin, alpha-actinin, myosin, and tropomyosin.
    • Employed fluorescein-labeled phalloidin to detect filamentous actin (F-actin).

    Main Results:

    • Observed the appearance of "flowers" on the dorsal cell surface within 15 minutes of temperature shift.
    • Identified significant accumulations of actin, alpha-actinin, myosin, and tropomyosin within these "flowers".
    • Demonstrated intense staining with phalloidin, indicating the presence of F-actin in these structures.

    Conclusions:

    • The early "flower" structures represent a rapid reorganization of microfilaments.
    • These findings suggest that microfilament dynamics are among the earliest events altered during RSV-induced cell transformation.

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