Background: Several actin nucleators, including Arp2/3 and various formins, control numerous cytoskeletal-based functions in vivo. Results: We investigated the relative roles of these nucleators. As a model system, we used natural killer (NK) lymphocytes, which display a wide range of cytoskeletal-based functions that culminate in the lysis of target cells. NK cells lacking either Arp2/3 or the formin hDia1 were ineffective in target cell lysis, but for distinct reasons. Loss of Arp2/3 function led to defects in cell adhesion and actin assembly at the junction with the target cell (the lytic synapse). In contrast, loss of hDia1 did not disrupt actin assembly at the lytic synapse. Instead, loss of hDia1 led to perturbations in the microtubule cytoskeleton, including the targeting of microtubules to the lytic synapse. Conclusions: These studies reveal novel distinctions and relationships among the functions of Arp2/3, formins, and microtubules in cells. Notably, a formin mediates the capture of microtubules at the cell periphery.