© The Rockefeller University Press,
0021-9525/2000//1075 $5.00
The Journal of Cell Biology, Volume 148, Number 5,
, 2000 1075-1090
Integrin Dynamics and Matrix Assembly
: Tensin-Dependent Translocation of
5β1 Integrins Promotes Early Fibronectin Fibrillogenesis
Roumen Pankova,
Edna Cukiermana,
Ben-Zion Katza,
Kazue Matsumotoa,
Diane C. Linb,
Shin Linb,
Cornelia Hahna, and
Kenneth M. Yamadaa
a Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892-4370
b Department of Developmental and Cell Biology, University of California, Irvine, Irvine, California 92697-1450
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.(301) 402-0897(301) 496-9124
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
vβ3 remains within focal contacts, the fibronectin receptor
5β1 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
5β1 integrins and depends on interactions with a functional actin cytoskeleton and vitronectin receptor ligation. During cell spreading, translocation of ligand-occupied
5β1 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
5β1 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
© 2000 The Rockefeller University Press
B.-Z. Katz's present address is The Hematology Institute, Tel-Aviv Medical Center, Weizman 6 Street, Tel-Aviv, Israel.
Abbreviations used in this paper: ECM, extracellular matrix; FC, focal contacts; FN, fibronectin; GFP, green fluorescent protein; HFF, human foreskin fibroblasts; VN, vitronectin.

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