Distinct developmental roles for direct and indirect talin-mediated linkage to actin.

Transmembrane adhesion receptors, such as integrins, mediate cell adhesion by interacting with intracellular proteins that connect to the cytoskeleton. Talin, one such linker protein, is essential to connect extracellular matrix-bound integrins to the cytoskeleton. Talin can connect to the cytoskeleton either directly, through its actin-binding motifs, or indirectly, by recruiting other actin-binding proteins. Talin's carboxy-terminal end contains a well-characterized actin-binding domain (ABD). We tested the role of the C-terminal ABD of talin in integrin function in Drosophila. We found that introduction of mutations that reduced actin binding in vitro into the isolated C-terminal Talin-ABD impaired actin binding in vivo. Moreover, when engineered into full-length talin, these mutations disrupted a subset of integrin-mediated adhesion-dependent developmental events. Specifically, morphogenetic processes that involve dynamic, short-term integrin-mediated adhesion were particularly sensitive to impaired function of the C-terminal Talin-ABD. We propose that during development talin connects integrins to the cytoskeleton in distinct ways in different types of integrin-mediated adhesion: directly in transient adhesions and indirectly in stable long-lasting adhesions. Our results provide insight into how a similar array of molecular components can contribute to diverse adhesive processes throughout development.

Crystal structure of the talin integrin binding domain 2.

Integrins are transmembrane receptors that mediate cell adhesion to the extracellular matrix and play essential roles in tissue development and maintenance. The cytoplasmic segment of integrin associates with talin, a large intracellular protein that links integrin to the actin cytoskeleton. Binding of talin via an integrin binding segment (IBS1) results in large conformational changes in the extracellular portion of integrin, which modulates the affinity of integrins for their extracellular matrix ligands. However, integrin binding also requires a second segment of talin (IBS2). Despite detailed descriptions of the integrin-IBS1 binding, the molecular determinants that drive the integrin-IBS2 association are poorly understood. Here, we describe the crystal structure of the talin IBS2 domain, which forms a five-helix bundle. The large structural homology with a vinculin binding domain hints at an ancient gene duplication and suggests that helix 4 may bind to vinculin if the bundle is unfolded. Mapping previous mutations on the surface highlights a likely binding interface for integrin.