Reduced expression of (alpha)5(beta)1 integrin prevents spreading-dependent cell proliferation.

Cell adhesion to substratum results in the initiation of integrin signaling and an integrin-dependent organization of the cytoskeleton (cell spreading). To address the potential relationships between these events and cell proliferation, we transfected NRK fibroblasts with an antisense cDNA encoding a 1.3 kb ATG-spanning portion of (alpha)5 integrin subunit and obtained stable clones in which the surface expression of (alpha)5(beta)1 integrin was selectively reduced. (alpha)5-antisense NRK cells are less spread than the control transfectants, have poorly defined stress fibers, and an increased amount of cortical actin. The antisense clones remained anchorage-dependent, but they proliferated very slowly. Serum dose-response curves showed that they have an impaired response to mitogens. Importantly, cell spreading and stress fiber formation could be completely restored by plating the antisense cells on collagen, but cell spreading failed to rescue proliferation. These data indicate that cell spreading can be uncoupled from cell proliferation and that cytoskeletal organization is important for NRK cell proliferation because it enforces the proliferative effect of (alpha)5(beta)1 integrin. Our results also indicate that reduced surface expression of (alpha)5(beta)1 integrin is not sufficient to confer the anchorage-independent phenotype to nontransformed cells.