Vinculin and talin: kinetics of entry and exit from the cytoskeletal pool.

Vinculin and talin, two major components of focal contacts, exist in cytosolic and cytoskeletal pools. The kinetics of entry and exit of the two proteins between the two pools were investigated in normal and transformed cells. In cultured chick embryo fibroblasts, a fraction (2-5%) of the newly synthesized vinculin and talin reached maximal levels in the cytoskeleton in 30-45 min. Both proteins had 2-3 times shorter half-lives in the cytoskeletal pool (t1/2 = 6-7 h) than in the cytosolic pool (t1/2 = 14-15 h), which suggests that the incorporation of cytosolic vinculin and talin into the cytoskeleton does not involve a simple equilibrium between the two pools. However, after disruption of cell-to-substrate adhesion by trypsinization, an equilibrium in the incorporation between the two pools was transiently established, resulting in the use of the preexisting cytosolic pools of the two proteins during re-establishment of cell-to-matrix contacts. Viral transformation did not cause a significant change in the incorporation rates into the cytoskeleton. However, it decreased the half-lives of both proteins in the cytoskeletal pool (t1/2 = approximately 4 h) and in the cytosolic pool (t1/2 = 9-10 h). The increased turn-over rates of vinculin and talin in the cytoskeletal pool in transformed cells may contribute to the enhanced motility of transformed cells.