Integrin-mediated cell adhesion transduces signaling activities for actin reorganization, which is crucially involved in cellular function and architectural integrity. In this study, we explored the possibility of whether cell-cell contacts might be regulated via integrin-α5β1-mediated actin reorganization. Ectopic expression of integrin α5 in integrin-α5-null intestinal epithelial cells resulted in facilitated retraction, cell-cell contact loss, and wound healing depending on Src and PI3K (phosphoinositide 3-kinase) activities by a reagent that affects actin organization. However, cytoplasmic tailless integrin α5 (hereafter referred to as α5/1) expression caused no such effects but rather sustained peripheral actin fibers, regardless of Src and PI3K signaling activities. Furthermore, integrin α5 engagement with fibronectin phosphorylated Ser643 of PKCδ, upstream of FAK and Src and at a transmodulatory loop with PI3K/Akt. Pharmacological PKCδ inactivation, dominant-negative PKCδ adenovirus or inactive cofilin phosphatase (SSH1L mutant) retrovirus infection of α5-expressing cells sustained peripheral actin organization and blocked the actin reorganizing-mediated loss of cell-cell contacts. Meanwhile, wild-type PKCδ expression sensitized α5/1-expressing cells to the actin disruptor to induce cell scattering. Altogether, these observations indicate that integrin α5, but not α5/1, mediates PKCδ phosphorylation and cofilin dephosphorylation, which in turn modulate peripheral actin organization presumably leading to an efficient regulation of cell-cell contact and migration.