Epidermal growth factor induces tyrosine phosphorylation and reorganization of the tight junction protein ZO-1 in A431 cells.
Addition of epidermal growth factor (EGF) to A431 human epidermal carcinoma cells results in actin reorganization and phosphorylation of several cytoskeletal proteins. In the present study, we found that EGF treatment of this cell line also results in the redistribution and tyrosine phosphorylation of ZO-1. In normal polarized epithelial cells, ZO-1 is restricted to the cytoplasmic surface of the most apical of the intercellular junctions, the tight junction. In contrast, ZO-1 in the majority of unstimulated A431 cells in small subconfluent islands colocalizes with actin along the lateral cell membranes and in rare microspikes and membrane ruffles. Exposure to EGF results in a transient redistribution of actin into an apically positioned ring. ZO-1 becomes highly focused at apical sites of cell contact and co-localizes with the newly formed band of perijunctional actin. Coincidently, ZO-1 and another tight junction protein, ZO-2, become transiently phosphorylated on tyrosine residues, as determined by anti-phosphotyrosine immunoblotting. Pre-treatment of A431 cells with cytochalasin D, which disrupts normal microfilament organization, does not affect EGF-dependent phosphorylation of the EGF receptor. However, cytochalasin D pretreatment blocks both the EGF-induced ZO-1 rearrangement and tyrosine phosphorylation, suggesting that these responses are dependent on an intact actin microfilament system. We speculate that the transient tyrosine phosphorylation of ZO-1 in response to EGF treatment may be involved in remodeling of intercellular junctions in A431 cells.
Van Itallie
C M
CM
Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520-8019, USA.
Balda
M S
MS
Anderson
J M
JM
D000199:Actins / Q000032:analysis / Q000187:drug effects / Q000378:metabolism; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell; D002460:Cell Line; D015638:Cytochalasin D / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D006801:Humans; D007365:Intercellular Junctions / Q000502:physiology / Q000648:ultrastructure; D008565:Membrane Proteins / Q000032:analysis / Q000187:drug effects / Q000378:metabolism*; D010750:Phosphoproteins / Q000032:analysis / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D062826:Zonula Occludens-1 Protein
D000199:Actins; D008565:Membrane Proteins; D010750:Phosphoproteins; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D019000:Phosphotyrosine; D015638:Cytochalasin D; D014443:Tyrosine; D004815:Epidermal Growth Factor