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© The Rockefeller University Press, 0021-9525/1999/11/879/ $5.00
The Journal of Cell Biology, Volume 147, Number 4, November 15, 1999 879-890

Original Article

Yes-associated Protein 65 Localizes p62c-Yes to the Apical Compartment of Airway Epithelia by Association with EBP50

Peter J. Mohlera, Silvia M. Kredab, Richard C. Boucherb, Marius Sudolc, M. Jackson Stuttsb, and Sharon L. Milgrama
a Department of Cell and Molecular Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599
b The Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599
c Department of Biochemistry, Mount Sinai Medical School, New York University, New York, NY 10029

Correspondence to: Sharon L. Milgram, Cell and Molecular Physiology, University of North Carolina at Chapel Hill, CB7545, Chapel Hill, NC 27599. Tel:(919) 966-9792 Fax:(919) 966-6927 E-mail:milg{at}med.unc.edu.

We recently showed that the COOH terminus of the cystic fibrosis transmembrane conductance regulator associates with the submembranous scaffolding protein EBP50 (ERM-binding phosphoprotein 50 kD; also called Na+/H+ exchanger regulatory factor). Since EBP50 associates with ezrin, this interaction links the cystic fibrosis transmembrane conductance regulator (CFTR) to the cortical actin cytoskeleton. EBP50 has two PDZ domains, and CFTR binds with high affinity to the first PDZ domain. Here, we report that Yes-associated protein 65 (YAP65) binds with high affinity to the second EBP50 PDZ domain. YAP65 is concentrated at the apical membrane in airway epithelia and interacts with EBP50 in cells. The COOH terminus of YAP65 is necessary and sufficient to mediate association with EBP50. The EBP50–YAP65 interaction is involved in the compartmentalization of YAP65 at the apical membrane since mutant YAP65 proteins lacking the EBP50 interaction motif are mislocalized when expressed in airway epithelial cells. In addition, we show that the nonreceptor tyrosine kinase c-Yes is contained within EBP50 protein complexes by association with YAP65. Subapical EBP50 protein complexes, containing the nonreceptor tyrosine kinase c-Yes, may regulate apical signal transduction pathways leading to changes in ion transport, cytoskeletal organization, or gene expression in epithelial cells.

Key Words: EBP50, NHERF, YAP65, Src family kinase, scaffolding protein


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