Yes-Associated Protein 65 Localizes P62c-Yes to the Apical Compartment of Airway Epithelia by Association with Ebp50

doi:10.1083/jcb.147.4.879

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

Original Article

Yes-Associated Protein 65 Localizes P62^c-Yes to the Apical Compartment of Airway Epithelia by Association with Ebp50

Peter J. Mohler^a, Silvia M. Kreda^b, Richard C. Boucher^b, Marius Sudol^c, M. Jackson Stutts^b, and Sharon L. Milgram^a

^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
Cell and Molecular Physiology, University of North Carolina at Chapel Hill, CB7545, Chapel Hill, NC 27599.(919) 966-6927(919) 966-9792

milg{at}med.unc.edu

We recently showed that the COOH terminus of the cystic fibrosistransmembrane conductance regulator associates with the submembranousscaffolding protein EBP50 (ERM-binding phosphoprotein 50 kD;also called Na⁺/H⁺ exchanger regulatory factor). Since EBP50associates with ezrin, this interaction links the cystic fibrosistransmembrane conductance regulator (CFTR) to the cortical actincytoskeleton. EBP50 has two PDZ domains, and CFTR binds withhigh affinity to the first PDZ domain. Here, we report thatYes-associated protein 65 (YAP65) binds with high affinity tothe second EBP50 PDZ domain. YAP65 is concentrated at the apicalmembrane in airway epithelia and interacts with EBP50 in cells.The COOH terminus of YAP65 is necessary and sufficient to mediateassociation with EBP50. The EBP50–YAP65 interaction isinvolved in the compartmentalization of YAP65 at the apicalmembrane since mutant YAP65 proteins lacking the EBP50 interactionmotif are mislocalized when expressed in airway epithelial cells.In addition, we show that the nonreceptor tyrosine kinase c-Yesis contained within EBP50 protein complexes by association withYAP65. Subapical EBP50 protein complexes, containing the nonreceptortyrosine kinase c-Yes, may regulate apical signal transductionpathways leading to changes in ion transport, cytoskeletal organization,or gene expression in epithelial cells.

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

Abbreviations used in this paper: CFTR, cystic fibrosis transmembraneconductance regulator; EBP50, ERM-binding phosphoprotein 50kD; E3KARP, NHE3 kinase A regulatory protein; ERM, ezrin-radixin-moesin;NHERF, Na⁺/H⁺ exchanger regulatory factor; NRTK, nonreceptortyrosine kinase; PKA, protein kinase A.

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