Fibronectin fragmentation promotes alpha4beta1 integrin-mediated contraction of a fibrin-fibronectin provisional matrix.
In injured tissues, the fibrin-fibronectin (FN) provisional matrix provides a framework for cell adhesion, migration, and repair. Effective repair and remodeling require a proper balance between extracellular matrix (ECM) deposition, contraction, and turnover. We utilized a three-dimensional (3D) fibrin-FN provisional matrix model to determine the contributions of the FN-binding integrin receptors alpha5beta1 and alpha4beta1 to matrix contraction. CHOalpha5 cells expressing alpha5beta1, a receptor for FN's RGD cell-binding domain, were highly contractile, and cells were well spread on a 3D fibrin-FN matrix. In contrast, CHOalpha4 cells expressing the alpha4beta1 receptor for FN's alternatively spliced V region attached less efficiently to FN and were deficient in fibrin-FN matrix contraction. Surprisingly, cell adhesion and matrix contraction by CHOalpha4 cells were dramatically enhanced, to levels equivalent to CHOalpha5 cells, when proteolyzed FN was used in place of intact FN in the fibrin-FN matrix. Similar enhancement was observed when ligand binding by alpha4beta1 integrins was activated by treatment with Mn(++), but not by stimulation of actin organization with LPA. Therefore, alpha4beta1-dependent cell responses to the provisional matrix are modulated by cleavage of matrix components.
Department of Molecular Biology, Princeton University, Princeton, NJ 08544-1014, USA.
Leyla V
LV
Valenick
Henry C
HC
Hsia
Jean E
JE
Schwarzbauer
D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002449:Cell Aggregation; D006224:Cricetinae; D005109:Extracellular Matrix / Q000502:physiology*; D005337:Fibrin / Q000502:physiology*; D005353:Fibronectins / Q000737:chemistry / Q000502:physiology*; D022001:Focal Adhesions / Q000502:physiology; D006801:Humans; D006868:Hydrolysis; D039041:Integrin alpha4beta1 / Q000502:physiology*; D010446:Peptide Fragments / Q000502:physiology; D011485:Protein Binding; D017469:Receptors, Fibronectin / Q000502:physiology; D011994:Recombinant Proteins
D005353:Fibronectins; D039041:Integrin alpha4beta1; D010446:Peptide Fragments; D017469:Receptors, Fibronectin; D011994:Recombinant Proteins; D005337:Fibrin