title: Alternative mechanisms for talin to mediate integrin function.
abstract: Cell-matrix adhesion is essential for building animals, promoting tissue cohesion, and enabling cells to migrate and resist mechanical force. Talin is an intracellular protein that is critical for linking integrin extracellular-matrix receptors to the actin cytoskeleton. A key question raised by structure-function studies is whether talin, which is critical for all integrin-mediated adhesion, acts in the same way in every context. We show that distinct combinations of talin domains are required for each of three different integrin functions during Drosophila development. The partial function of some mutant talins requires vinculin, indicating that recruitment of vinculin allows talin to duplicate its own activities. The different requirements are best explained by alternative mechanisms of talin function, with talin using one or both of its integrin-binding sites. We confirmed these alternatives by showing that the proximity between the second integrin-binding site and integrins differs, suggesting that talin adopts different orientations relative to integrins. Finally, we show that vinculin and actomyosin activity help change talin's orientation. These findings demonstrate that the mechanism of talin function differs in each developmental context examined. The different arrangements of the talin molecule relative to integrins suggest that talin is able to sense different force vectors, either parallel or perpendicular to the membrane. This provides a paradigm for proteins whose apparent uniform function is in fact achieved by a variety of distinct mechanisms involving different molecular architectures.
authors.affiliation: The Gurdon Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Klapholz
authors.affiliation: The Gurdon Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Samantha L
authors.initials: SL
authors.lastname: Herbert
authors.affiliation: The Gurdon Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Jutta
authors.initials: J
authors.lastname: Wellmann
authors.affiliation: The Gurdon Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
authors.forename: Robert
authors.initials: R
authors.lastname: Johnson
authors.affiliation: Randall Division of Cell and Molecular Biophysics, King's College London, London SE1 1UL, UK.
authors.forename: Maddy
authors.initials: M
authors.lastname: Parsons
authors.affiliation: The Gurdon Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK. Electronic address: nb117@cam.ac.uk.
authors.forename: Nicholas H
authors.initials: NH
authors.lastname: Brown
mesh_terms: D000205:Actomyosin / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology*; D004330:Drosophila / Q000254:growth & development*; D005109:Extracellular Matrix / Q000378:metabolism*; D016023:Integrins / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D016608:Talin / Q000235:genetics / Q000378:metabolism*; D016596:Vinculin / Q000378:metabolism
chemical_list: D016023:Integrins; D016608:Talin; D016596:Vinculin; D000205:Actomyosin