The aggregation of the high affinity IgE receptor induces tyrosine phosphorylation of paxillin, a focal adhesion protein.
Tyrosine phosphorylation of proteins is an essential component of high affinity IgE receptor (Fc epsilon RI) signaling and secretion. This signaling and secretion is also dependent on the organization of the cytoskeleton. Here we report that the aggregation of Fc epsilon RI on rat basophilic leukemia cells results in tyrosine phosphorylation of the cytoskeletal protein, paxillin. Tyrosine phosphorylation of paxillin is a relatively late event after Fc epsilon RI aggregation. Both the direct increase in intracellular Ca2+ with calcium ionophore and the activation of protein kinase C (PKC) with PMA induced tyrosine phosphorylation of paxillin. The optimal tyrosine phosphorylation of paxillin by Fc epsilon RI aggregation required PKC and extracellular Ca2+. However, there was also Fc epsilon RI-mediated tyrosine phosphorylation of paxillin independent of Ca2+ influx or PKC activation. By fluorescent microscopy, cell stimulation induced a redistribution of paxillin toward the periphery of the cells. Although Fc epsilon RI aggregation induced tyrosine phosphorylation of paxillin in nonadherent cells, adherence markedly enhanced this phosphorylation. Together, the data suggest a role for paxillin in Fc epsilon RI signaling.
Laboratory of Immunology, National Institute of Dental Research, National Institutes of Health, Bethesda, MD 20892.
M M
MM
Hamawy
W D
WD
Swaim
K
K
Minoguchi
A W
AW
de Feijter
S E
SE
Mergenhagen
R P
RP
Siraganian
D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion / Q000276:immunology; D003598:Cytoskeletal Proteins / Q000276:immunology / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods; D051419:Paxillin; D010750:Phosphoproteins / Q000276:immunology / Q000378:metabolism*; D011233:Precipitin Tests; D011493:Protein Kinase C / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats; D011940:Receptor Aggregation / Q000276:immunology*; D017455:Receptors, IgE / Q000276:immunology*; D014407:Tumor Cells, Cultured
D003598:Cytoskeletal Proteins; D051419:Paxillin; D010750:Phosphoproteins; C496063:Pxn protein, rat; D017455:Receptors, IgE; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C; D002118:Calcium