title: Vinculin controls talin engagement with the actomyosin machinery.
abstract: The link between extracellular-matrix-bound integrins and intracellular F-actin is essential for cell spreading and migration. Here, we demonstrate how the actin-binding proteins talin and vinculin cooperate to provide this link. By expressing structure-based talin mutants in talin null cells, we show that while the C-terminal actin-binding site (ABS3) in talin is required for adhesion complex assembly, the central ABS2 is essential for focal adhesion (FA) maturation. Thus, although ABS2 mutants support cell spreading, the cells lack FAs, fail to polarize and exert reduced force on the surrounding matrix. ABS2 is inhibited by the preceding mechanosensitive vinculin-binding R3 domain, and deletion of R2R3 or expression of constitutively active vinculin generates stable force-independent FAs, although cell polarity is compromised. Our data suggest a model whereby force acting on integrin-talin complexes via ABS3 promotes R3 unfolding and vinculin binding, activating ABS2 and locking talin into an actin-binding configuration that stabilizes FAs.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Atherton
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Ben
authors.initials: B
authors.lastname: Stutchbury
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, UK.
authors.forename: De-Yao
authors.initials: DY
authors.lastname: Wang
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Devina
authors.initials: D
authors.lastname: Jethwa
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Ricky
authors.initials: R
authors.lastname: Tsang
authors.affiliation: Department of Microbiology, Complutense, University of Madrid, 28040 Madrid, Spain.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Meiler-Rodriguez
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Pengbo
authors.initials: P
authors.lastname: Wang
authors.affiliation: Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: Neil
authors.initials: N
authors.lastname: Bate
authors.affiliation: Vanderbilt Centre for Kidney Disease, Vanderbilt Division of Nephrology, S-3223 Medical Centre, North Nashville, Tennessee, USA.
authors.forename: Roy
authors.initials: R
authors.lastname: Zent
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Crown Street, Liverpool L69 7ZB, UK.
authors.forename: Igor L
authors.initials: IL
authors.lastname: Barsukov
authors.affiliation: School of Biosciences, University of Kent, Canterbury, KENT CT2 7NJ, UK.
authors.forename: Benjamin T
authors.initials: BT
authors.lastname: Goult
authors.affiliation: Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: David R
authors.initials: DR
authors.lastname: Critchley
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Christoph
authors.initials: C
authors.lastname: Ballestrem
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000199:Actins / Q000235:genetics; D000205:Actomyosin / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D016764:Cell Polarity; D022001:Focal Adhesions / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016608:Talin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016596:Vinculin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D016608:Talin; D016596:Vinculin; D000205:Actomyosin