Association of beta 1 integrin with focal adhesion kinase and paxillin in differentiating Schwann cells.
Schwann cells (SCs) differentiate into a myelinating cell when simultaneously adhering to an axon destined for myelination and basal lamina. We are interested in defining the signaling pathway activated by basal lamina. Using SC/sensory neuron (N) cocultures, we identified beta1 integrin and F-actin as components of a pathway leading to myelin gene expression and myelination (Fernandez-Valle et al., 1994, 1997). Here, we show that focal adhesion kinase (FAK) and paxillin are constitutively expressed by SCs contacting axons in the absence of basal lamina. Tyrosine phosphorylation of FAK and paxillin increases as SCs form basal lamina and differentiate. FAK and paxillin specifically coimmunoprecipitate with beta1 integrin in differentiating SC/N cocultures but not SC-only cultures. Paxillin coimmunoprecipitates with FAK and fyn kinase in differentiating SC/N cocultures. A subset of tyrosine-phosphorylated beta1 integrin, FAK, and paxillin molecules reside in the insoluble, F-actin-rich fraction of differentiating cocultures. Cytochalasin D, an actin depolymerizing agent, decreases tyrosine phosphorylation of FAK and paxillin and their association with beta1 integrin and causes a dose-dependent increase in the abundance of insoluble FAK and paxillin complexes. Collectively, our work indicates that beta1 integrin, FAK, paxillin, and fyn kinase form an actin-associated complex in SCs adhering to basal lamina in the presence of axons. This complex may be important for initiating the process of SC differentiation into a myelinating cell.
Chen
L M
LM
Department of Molecular Biology and Microbiology, University of Central Florida, Orlando, Florida 32816-2360, USA.
Bailey
D
D
Fernandez-Valle
C
C
D000818:Animals; D000831:Animals, Newborn; D000906:Antibodies; D001369:Axons / Q000737:chemistry / Q000201:enzymology; D002450:Cell Communication / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D015638:Cytochalasin D / Q000494:pharmacology; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D003599:Cytoskeleton / Q000737:chemistry; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019012:Integrin beta1 / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D009475:Neurons, Afferent / Q000166:cytology / Q000648:ultrastructure; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology*; D015398:Signal Transduction / Q000502:physiology; D014443:Tyrosine / Q000378:metabolism
D000906:Antibodies; D003598:Cytoskeletal Proteins; D019012:Integrin beta1; D019384:Nucleic Acid Synthesis Inhibitors; D051419:Paxillin; D010750:Phosphoproteins; C496063:Pxn protein, rat; D015638:Cytochalasin D; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496052:Ptk2 protein, rat