Original Article

Integrin Dynamics and Matrix Assembly: Tensin-dependent Translocation of ₅ß₁ Integrins Promotes Early Fibronectin Fibrillogenesis

Correspondence to: Kenneth M. Yamada, Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Building 30, Room 421, 30 Convent Drive MSC 4370, Bethesda, MD 20892-4370. Tel:(301) 496-9124 Fax:(301) 402-0897 E-mail:ky4w{at}nih.gov.

Fibronectin matrix assembly is a multistep, integrin-dependent process. To investigate the role of integrin dynamics in fibronectin fibrillogenesis, we developed an antibody-chasing technique for simultaneous tracking of two integrin populations by different antibodies. We established that whereas the vitronectin receptor _vß₃ remains within focal contacts, the fibronectin receptor ₅ß₁ translocates from focal contacts into and along extracellular matrix (ECM) contacts. This escalator-like translocation occurs relative to the focal contacts at 6.5 ± 0.7 µm/h and is independent of cell migration. It is induced by ligation of ₅ß₁ integrins and depends on interactions with a functional actin cytoskeleton and vitronectin receptor ligation. During cell spreading, translocation of ligand-occupied ₅ß₁ integrins away from focal contacts and along bundles of actin filaments generates ECM contacts. Tensin is a primary cytoskeletal component of these ECM contacts, and a novel dominant-negative inhibitor of tensin blocked ECM contact formation, integrin translocation, and fibronectin fibrillogenesis without affecting focal contacts. We propose that translocating ₅ß₁ integrins induce initial fibronectin fibrillogenesis by transmitting cytoskeleton-generated tension to extracellular fibronectin molecules. Blocking this integrin translocation by a variety of treatments prevents the formation of ECM contacts and fibronectin fibrillogenesis. These studies identify a localized, directional, integrin translocation mechanism for matrix assembly.

Key Words: fibronectin, integrin, tensin, vitronectin, extracellular matrix

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