Ezrin directly interacts with AQP2 and promotes its endocytosis

Wei Li, William W. Jin, Kenji Tsuji, Ying Chen, Naohiro Nomura, Limin Su, Naofumi Yui, Julian Arthur, Susanna Cotecchia, Teodor G. Păunescu, Dennis Brown, Hua A.J. Lu

Research output: Contribution to journalArticle

9 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2914-2925
Number of pages12
JournalJournal of Cell Science
Volume130
Issue number17
DOIs
Publication statusPublished - Sep 1 2017
Externally publishedYes

Fingerprint

Aquaporin 2
Endocytosis
Vasopressins
Actin Cytoskeleton
ezrin
Microfilament Proteins
Aquaporins
Membranes
Protein Transport
Recombinant Proteins
Proteomics
Small Interfering RNA
Fluorescent Antibody Technique

Keywords

  • Aquaporin-2
  • Endocytosis
  • Ezrin
  • Trafficking
  • Water homeostasis

ASJC Scopus subject areas

  • Cell Biology

Cite this

Li, W., Jin, W. W., Tsuji, K., Chen, Y., Nomura, N., Su, L., ... Lu, H. A. J. (2017). Ezrin directly interacts with AQP2 and promotes its endocytosis. Journal of Cell Science, 130(17), 2914-2925. https://doi.org/10.1242/jcs.204842

Ezrin directly interacts with AQP2 and promotes its endocytosis. / Li, Wei; Jin, William W.; Tsuji, Kenji; Chen, Ying; Nomura, Naohiro; Su, Limin; Yui, Naofumi; Arthur, Julian; Cotecchia, Susanna; Păunescu, Teodor G.; Brown, Dennis; Lu, Hua A.J.

In: Journal of Cell Science, Vol. 130, No. 17, 01.09.2017, p. 2914-2925.

Research output: Contribution to journalArticle

Li, W, Jin, WW, Tsuji, K, Chen, Y, Nomura, N, Su, L, Yui, N, Arthur, J, Cotecchia, S, Păunescu, TG, Brown, D & Lu, HAJ 2017, 'Ezrin directly interacts with AQP2 and promotes its endocytosis', Journal of Cell Science, vol. 130, no. 17, pp. 2914-2925. https://doi.org/10.1242/jcs.204842
Li, Wei ; Jin, William W. ; Tsuji, Kenji ; Chen, Ying ; Nomura, Naohiro ; Su, Limin ; Yui, Naofumi ; Arthur, Julian ; Cotecchia, Susanna ; Păunescu, Teodor G. ; Brown, Dennis ; Lu, Hua A.J. / Ezrin directly interacts with AQP2 and promotes its endocytosis. In: Journal of Cell Science. 2017 ; Vol. 130, No. 17. pp. 2914-2925.
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