Actin cytoskeleton polymerization in Dbl-transformed NIH3T3 fibroblasts is dependent on cell adhesion to specific extracellular matrix proteins.
The Dbl oncogene is the putative exchange factor for two small GTP-binding proteins, RhoA and CDC42 which are involved in the polymerization of actin to produce stress fibers and filopodia, respectively. We report here that Dbl oncogene-transformed NIH3T3 cells show actin stress fibers only when cells are plated on fibronectin. Plating of cells on collagen I and IV as well as on poly-D-lysine and gelatin induces polymerization of actin to form filopodia, lamellipodia and membrane ruffles but not stress fibers. The putative collagen receptors, alpha1/beta1 and alpha2/beta1 integrins are expressed at reduced level in Dbl-transformed cells compared to untransformed NIH3T3 fibroblasts. Nevertheless, adhesion to collagens is not altered. Inhibitory monoclonal antibody to mouse integrin beta1 subunit blocked adhesion of both Dbl-transformed and untransformed NIH3T3 cells, demonstrating that adhesion to collagen I and IV is mediated by the beta1 family of integrins. Dbl product rapidly induces the depolymerization of actin stress fibers, rounding up of the cells, and formation of filopodia and lamellipodia when microinjected in NIH3T3 cells plated on gelatin. Thus, Dbl may exert its effect on actin cytoskeleton organization in response to extracellular proteins by altering integrin-mediated signalling pathways.
Defilippi
P
P
Dipartimento di Genetica, Biologia e Chimica Medica, Universita di Torino, Italy.
Olivo
C
C
Tarone
G
G
Mancini
P
P
Torrisi
M R
MR
Eva
A
A
D016475:3T3 Cells; D000199:Actins / Q000502:physiology*; D000818:Animals; D002448:Cell Adhesion; D002455:Cell Division; D002461:Cell Line, Transformed; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D005109:Extracellular Matrix; D020662:Guanine Nucleotide Exchange Factors; D016023:Integrins / Q000096:biosynthesis; D051379:Mice; D008845:Microinjections; D009857:Oncogenes; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000494:pharmacology; D015682:Retroviridae Proteins, Oncogenic / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D014162:Transfection
D000199:Actins; D020662:Guanine Nucleotide Exchange Factors; D016023:Integrins; C512843:Mcf2 protein, mouse; D011993:Recombinant Fusion Proteins; D015682:Retroviridae Proteins, Oncogenic