Elsevier

Experimental Cell Research

Volume 173, Issue 1, November 1987, Pages 244-255
Experimental Cell Research

Original article
Organization of stress fibers in cultured fibroblasts after extraction of actin with bovine brain gelsolin-like protein

https://doi.org/10.1016/0014-4827(87)90349-1Get rights and content

Abstract

Mouse and quail embryo fibroblasts were extracted with Triton X-100 and the resulting cytoskeletons were treated with gelsolin-like actin-capping protein (the 90-kDa protein-actin complex isolated from bovine brain). Staining of cells with rhodamine-conjugated phalloin or an antibody to actin did not reveal any actin-containing structures after treatment with the 90-kDa protein-actin complex. Extraction of actin was confirmed by SDS-gel electrophoresis. Immunofluorescence microscopy showed that vinculin and α-actinin were released from the cytoskeletons together with actin. However, myosin remained associated with the cytoskeleton after treatment with the 90-kDa protein-actin complex. The distribution of myosin in treated cells showed no significant difference from that in control cells: in both cases myosin was localized mainly in the stress fibers. Double-fluorescence staining showed the absence of actin in myosin-containing stress fibers of treated cells. The ultrastructural organization of actin-depleted stress fibers was studied by transmission electron microscopy of platinum replicas. On electron micrographs these fibers appeared as bundles of filaments containing clusters of globular material. It is concluded that myosin localization in stress fibers does not depend on actin.

References (18)

  • R. Karlsson et al.

    Exp. Cell Res

    (1985)
  • A.B. Verkhovsky et al.

    Biochem. Biophys. Res. Commun

    (1984)
  • A.A. Neyfakh et al.

    Exp. Cell Res

    (1983)
  • A.D. Bershadsky et al.

    Exp. Cell Res

    (1985)
  • A.D. Bershadsky et al.

    Cell Biol. Int. Rep

    (1978)
  • H.R. Byers et al.
  • P. Mangeat et al.

    J. Cell Biol

    (1984)
  • G. Langanger et al.

    J. Cell Biol

    (1986)
  • Z. Avnur et al.

    J. Cell Biol

    (1983)
There are more references available in the full text version of this article.

Cited by (21)

View all citing articles on Scopus
View full text