Hepatocyte growth factor alters the polarity of Madin-Darby canine kidney cell monolayers.
Hepatocyte growth factor (HGF) and E-cadherin are important for epithelial morphogenetic events. We examined the effects of HGF on E-cadherin localization and interaction with beta-catenin in polarized Madin-Darby canine kidney (MDCK) cell monolayers grown on filters. Surface biotinylation experiments showed that HGF increases apically accessible E-cadherin. Confocal immunofluorescence microscopy of HGF-treated cells showed localization of E-cadherin at membrane domains contacting the apical compartment and an increase in accessibility of apically applied antibodies to lateral E-cadherin below the tight junction. Coimmunoprecipitation of beta-catenin/E-cadherin complexes showed that the amount of E-cadherin associated with beta-catenin increased during the first 24 h of HGF treatment with a return to baseline values after 48 and 72 h. Metabolic labeling showed that HGF increased the synthetic rate of beta-catenin and the amount of newly synthesized E-cadherin associated with immunoprecipitated beta-catenin, with the peak effect occurring after 12 h of treatment and returning to baseline after 24 h. HGF treatment inhibited transcytosis of immunoglobulin A by the polymeric immunoglobulin receptor. We conclude that HGF treatment of polarized MDCK cells grown on filters decreases cell polarity and alters E-cadherin/beta-catenin interaction and synthesis.
Balkovetz
D F
DF
Department of Anatomy, Cardiovascular Research Institute, University of California, San Francisco, California 94143-0452, USA. balkovetz@nrtc.dom.uab.edu
Pollack
A L
AL
Mostov
K E
KE
D000818:Animals; D001710:Biotin / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D016764:Cell Polarity / Q000187:drug effects*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004285:Dogs; D019084:Fluorescent Antibody Technique, Indirect; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D007070:Immunoglobulin A / Q000378:metabolism; D007668:Kidney / Q000187:drug effects*; D018613:Microscopy, Confocal; D051178:alpha Catenin
D015820:Cadherins; D003598:Cytoskeletal Proteins; D007070:Immunoglobulin A; D051178:alpha Catenin; D017228:Hepatocyte Growth Factor; D001710:Biotin