title: Proteolytic activity of specialized surface protrusions formed at rosette contact sites of transformed cells.
abstract: Surface protrusions at the leading edge of a moving cell that make contact with the surrounding extracellular matrix (ECM) are its main motor for locomotion and invasion. Chicken embryonic fibroblasts transformed by Rous sarcoma virus (RSV-CEF) form specialized membrane rosette-shaped contact sites on planar substrata as shown by interference reflection microscopy (IRM). Such activity is lacking in normal cells. These rosette contacts are more labile than other adhesion sites, such as focal and close contacts. Ultrastructural studies demonstrate that rosettes are sites at which membrane protrusions from the ventral cell surface contact the substratum. These protrusions are filled with meshworks of microfilaments and contain the pp60src oncogene product, actin, vinculin, and alpha-actinin. However, unlike focal contacts, at the rosettes these proteins interact to extend a highly motile membrane. Rosettes have the biological activity of degrading ECM components, as demonstrated by (1) local degradation of fibronectin substrata at sites of rosette contacts, but not focal and close contacts; (2) localization of putative antiprotease antibody at sites of rosette contacts, but not at focal an close contacts; and (3) local disruption of fibronectin matrix at sites of protrusive activity seen by transmission electron microscopy (TEM). In addition, formation of the rosette contact is insensitive to the ionophore monensin, and to inhibitors of proteolytic enzymes, while local fibronectin degradation at rosette contacts is inhibited by inhibitors of metalloproteases, 1,10-phenanthroline and NP-20. I consider these membrane protrusions of the rosette contacts in RSV-transformed cells specialized structural entities--invadopodia--that are involved in the local degradation of the ECM.
authors.affiliation: Department of Anatomy and Cell Biology, Georgetown University Medical School, Washington, D.C. 20007.
authors.forename: W T
authors.initials: WT
authors.lastname: Chen
mesh_terms: D000818:Animals; D001358:Avian Sarcoma Viruses; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure*; D002465:Cell Movement; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D005780:Gelatin; D006801:Humans; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D015688:Oncogene Protein pp60(v-src); D010447:Peptide Hydrolases / Q000378:metabolism; D010784:Photomicrography; D011480:Protease Inhibitors / Q000378:metabolism; D012191:Retroviridae Proteins / Q000378:metabolism; D016596:Vinculin
chemical_list: D003598:Cytoskeletal Proteins; D005353:Fibronectins; D011480:Protease Inhibitors; D012191:Retroviridae Proteins; D016596:Vinculin; D005780:Gelatin; D015688:Oncogene Protein pp60(v-src); D010447:Peptide Hydrolases