title: The beta1-integrin cytosolic domain optimizes phospholipase A2-mediated arachidonic acid release required for NIH-3T3 cell spreading.
abstract: Inhibition of PLA2 activity and rescue by addition of exogenous AA was used to demonstrate that AA production is essential for integrin-mediated NIH-3T3 murine cell spreading. Both AA release and cell spreading after attachment to a FN substrate were inhibited by the PLA2 inhibitor mepacrine. AA release was essential for signaling spreading since the inhibition of spreading induced by mepacrine was overcome by exogenous AA. Cells ectopically expressing full-length chicken beta1-integrins both released AA and spread fully on a substrate of anti-chicken beta1-integrin monoclonal antibody, and inhibition of PLA2 by mepacrine suppressed both spreading and AA release. Exogenous AA also reversed this mepacrine-induced inhibition of spreading. The role of the beta1-integrin cytosolic domain in AA release was examined by comparing responses of cells expressing full-length chicken beta1-integrins versus cells expressing a deletion mutant chicken beta1-integrin with a truncated cytosolic domain. Cells expressing a truncated chicken beta1-integrin released significantly less AA and failed to spread on the anti-chicken beta1-integrin antibody substrate. Furthermore, clustering full-length receptors with soluble antibody stimulated greater AA release than clustering of receptors having truncated cytosolic domains. These data suggest the beta1-integrin cytosolic domain is required for optimal PLA2 activation to produce AA necessary for cell spreading.
mesh_terms: D016475:3T3 Cells; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D002465:Cell Movement; D002645:Chickens; D003600:Cytosol / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019012:Integrin beta1 / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054467:Phospholipases A2; D011796:Quinacrine / Q000494:pharmacology