@article{b08459ca3c2749d2b49a2f14e0d89a2d,
title = "Ezrin directly interacts with AQP2 and promotes its endocytosis",
abstract = "The water channel aquaporin-2 (AQP2) is a major regulator of water homeostasis in response to vasopressin (VP). Dynamic trafficking of AQP2 relies on its close interaction with trafficking machinery proteins and the actin cytoskeleton. Here, we report the identification of ezrin, an actin-binding protein from the ezrin/radixin/moesin (ERM) family as an AQP2-interacting protein. Ezrin was first detected in a coimmunoprecipitation (co-IP) complex using an anti-AQP2 antibody in a proteomic analysis. Immunofluorescence staining revealed the coexpression of ezrin and AQP2 in collecting duct principal cells, and VP treatment caused redistribution of both proteins to the apical membrane. The ezrin-AQP2 interaction was confirmed by co-IP experiments with an anti-ezrin antibody, and by pulldown assays using purified full-length and FERM domain-containing recombinant ezrin. By using purified recombinant proteins, we showed that ezrin directly interacts with AQP2 C-terminus through its N-terminal FERM domain. Knocking down ezrin expression with shRNA resulted in increased membrane accumulation of AQP2 and reduced AQP2 endocytosis. Therefore, through direct interaction with AQP2, ezrin facilitates AQP2 endocytosis, thus linking the dynamic actin cytoskeleton network with AQP2 trafficking.",
keywords = "Aquaporin-2, Endocytosis, Ezrin, Trafficking, Water homeostasis",
author = "Wei Li and Jin, {William W.} and Kenji Tsuji and Ying Chen and Naohiro Nomura and Limin Su and Naofumi Yui and Julian Arthur and Susanna Cotecchia and P{\u a}unescu, {Teodor G.} and Dennis Brown and Lu, {Hua A.J.}",
note = "Funding Information: The authors would like to thank Dr Quanhong Qi for her technical assistance and Dr Matthew J. Mahon for helpful discussions of the project. The authors also thank Dr Tomas Kirchhausen for the kind gift of ezrin shRNA plasmids. The Zeiss LSM800 Airyscan system was purchased using an NIH Shared Instrumentation Grant (S10 OD021577-01) to D.B. S.C. is supported by Telethon Foundation, Italy (GGP13227). Additional support for the Program in Membrane Biology Microscopy Core comes from the Boston Area Diabetes and Endocrinology Research Center (DK57521) and the MGH Center for the Study of Inflammatory Bowel Disease (DK43351). Funding Information: HAJL is supported by the National Institutes of Health (NIH) (R01DK096015 and R21DK092619), the NephCure Foundation, a Gottschalk research grant from the American Society of Nephrology, and the Massachusetts General Hospital Executive Committee on Research. D.B. is supported by the NIH (R01DK096586). Deposited in PMC for release after 12 months. Publisher Copyright: {\textcopyright} 2017. Published by The Company of Biologists Ltd.",
year = "2017",
month = sep,
day = "1",
doi = "10.1242/jcs.204842",
language = "English",
volume = "130",
pages = "2914--2925",
journal = "The Quarterly journal of microscopical science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "17",
}