Unphosphorylated gelsolin is localized in regions of cell-substratum contact or attachment in Rous sarcoma virus-transformed rat cells.
Regions associated with cell-substratum contact or attachment in Rous sarcoma virus (RSV)-transformed rat fibroblasts (RR1022 cells) were identified by reflection-interference microscopy. Electron microscopy of such regions revealed the presence of discrete membrane-associated structures composed of a paracrystalline lattice of hexagons and pentagons to which actin filaments appear to be attached. Staining of actin by biotin-labeled heavy meromyosin showed that transformed cells, unlike normal fibroblasts, lack prominent actin fibers, and that, instead, much of the fluorescence is concentrated in loci corresponding to locations of transient association between the cell and the substratum. In stationary cells, such loci were found in rosette formation, predominantly in the region beneath the nucleus. In cells engaged in active movement, such as during migration into a wound, the actin-containing spots were concentrated in the region of the leading edge. A similar pattern of staining was observed with antibody to gelsolin, a 91,000-dalton Ca2+-dependent actin filament-shortening protein. Since the action of gelsolin on actin is reversible and dependent on physiologically relevant changes in calcium concentration, the localization of gelsolin, together with actin-bundling proteins such as alpha-actinin, in the regions containing many small microfilament bundles on the ventral side of cytoplasm suggests that gelsolin may be a component of the mechanism for the disassembly and assembly of actin during the dissolution and reformation of structures for cell-substratum contact during cell locomotion. Regulation of gelsolin activity was not dependent on protein phosphorylation, as shown by lack of 32P-incorporation into gelsolin in either transformed or normal fibroblasts.
D000199:Actins; D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics*; D002135:Calcium-Binding Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement; D002471:Cell Transformation, Neoplastic; D003599:Cytoskeleton / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D018260:Gelsolin; D007668:Kidney; D008264:Macrophages / Q000032:analysis; D008854:Microscopy, Electron; D051381:Rats; D012513:Sarcoma, Experimental / Q000648:ultrastructure
D000199:Actins; D002135:Calcium-Binding Proteins; D018260:Gelsolin