PLEKHA7 Recruits PDZD11 to Adherens Junctions to Stabilize Nectins

Diego Guerrera; Jimit Shah; Ekaterina Vasileva; Sophie Sluysmans; Isabelle Méan; Lionel Jond; Ina Poser; Matthias Mann; Anthony A Hyman; Sandra Citi

doi:10.1074/jbc.m115.712935

PLEKHA7 Recruits PDZD11 to Adherens Junctions to Stabilize Nectins

Diego Guerrera, Jimit Shah, Ekaterina Vasileva, Sophie Sluysmans, Isabelle Méan, Lionel Jond, Ina Poser, Matthias Mann, Anthony A Hyman, Sandra Citi

13 Citationer (Scopus)

Abstract

PLEKHA7 is a junctional protein implicated in stabilization of the cadherin protein complex, hypertension, cardiac contractility, glaucoma, microRNA processing, and susceptibility to bacterial toxins. To gain insight into the molecular basis for the functions of PLEKHA7, we looked for new PLEKHA7 interactors. Here, we report the identification of PDZ domain-containing protein 11 (PDZD11) as a new interactor of PLEKHA7 by yeast two-hybrid screening and by mass spectrometry analysis of PLEKHA7 immunoprecipitates. We show that PDZD11 (17 kDa) is expressed in epithelial and endothelial cells, where it forms a complex with PLEKHA7, as determined by co-immunoprecipitation analysis. The N-terminal Trp-Trp (WW) domain of PLEKHA7 interacts directly with the N-terminal 44 amino acids of PDZD11, as shown by GST-pulldown assays. Immunofluorescence analysis shows that PDZD11 is localized at adherens junctions in a PLEKHA7-dependent manner, because its junctional localization is abolished by knock-out of PLEKHA7, and is rescued by re-expression of exogenous PLEKHA7. The junctional recruitment of nectin-1 and nectin-3 and their protein levels are decreased via proteasome-mediated degradation in epithelial cells where either PDZD11 or PLEKHA7 have been knocked-out. PDZD11 forms a complex with nectin-1 and nectin-3, and its PDZ domain interacts directly with the PDZ-binding motif of nectin-1. PDZD11 is required for the efficient assembly of apical junctions of epithelial cells at early time points in the calcium-switch model. These results show that the PLEKHA7-PDZD11 complex stabilizes nectins to promote efficient early junction assembly and uncover a new molecular mechanism through which PLEKHA7 recruits PDZ-binding membrane proteins to epithelial adherens junctions.

Originalsprog	Engelsk
Tidsskrift	The Journal of Biological Chemistry
Vol/bind	291
Udgave nummer	21
Sider (fra-til)	11016-29
Antal sider	14
ISSN	0021-9258
DOI	https://doi.org/10.1074/jbc.m115.712935
Status	Udgivet - 20 maj 2016
Udgivet eksternt	Ja

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10.1074/jbc.m115.712935

http://www.jbc.org/content/291/21/11016.full.pdfLicens: CC BY

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title = "PLEKHA7 Recruits PDZD11 to Adherens Junctions to Stabilize Nectins",

abstract = "PLEKHA7 is a junctional protein implicated in stabilization of the cadherin protein complex, hypertension, cardiac contractility, glaucoma, microRNA processing, and susceptibility to bacterial toxins. To gain insight into the molecular basis for the functions of PLEKHA7, we looked for new PLEKHA7 interactors. Here, we report the identification of PDZ domain-containing protein 11 (PDZD11) as a new interactor of PLEKHA7 by yeast two-hybrid screening and by mass spectrometry analysis of PLEKHA7 immunoprecipitates. We show that PDZD11 (17 kDa) is expressed in epithelial and endothelial cells, where it forms a complex with PLEKHA7, as determined by co-immunoprecipitation analysis. The N-terminal Trp-Trp (WW) domain of PLEKHA7 interacts directly with the N-terminal 44 amino acids of PDZD11, as shown by GST-pulldown assays. Immunofluorescence analysis shows that PDZD11 is localized at adherens junctions in a PLEKHA7-dependent manner, because its junctional localization is abolished by knock-out of PLEKHA7, and is rescued by re-expression of exogenous PLEKHA7. The junctional recruitment of nectin-1 and nectin-3 and their protein levels are decreased via proteasome-mediated degradation in epithelial cells where either PDZD11 or PLEKHA7 have been knocked-out. PDZD11 forms a complex with nectin-1 and nectin-3, and its PDZ domain interacts directly with the PDZ-binding motif of nectin-1. PDZD11 is required for the efficient assembly of apical junctions of epithelial cells at early time points in the calcium-switch model. These results show that the PLEKHA7-PDZD11 complex stabilizes nectins to promote efficient early junction assembly and uncover a new molecular mechanism through which PLEKHA7 recruits PDZ-binding membrane proteins to epithelial adherens junctions.",

keywords = "Adherens Junctions, Animals, Carrier Proteins, Cell Adhesion Molecules, Dogs, Humans, Madin Darby Canine Kidney Cells, Multiprotein Complexes, Nectins, Journal Article, Research Support, Non-U.S. Gov't",

author = "Diego Guerrera and Jimit Shah and Ekaterina Vasileva and Sophie Sluysmans and Isabelle M{\'e}an and Lionel Jond and Ina Poser and Matthias Mann and Hyman, {Anthony A} and Sandra Citi",

note = "{\textcopyright} 2016 by The American Society for Biochemistry and Molecular Biology, Inc.",

year = "2016",

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doi = "10.1074/jbc.m115.712935",

language = "English",

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T1 - PLEKHA7 Recruits PDZD11 to Adherens Junctions to Stabilize Nectins

AU - Guerrera, Diego

AU - Shah, Jimit

AU - Vasileva, Ekaterina

AU - Sluysmans, Sophie

AU - Méan, Isabelle

AU - Jond, Lionel

AU - Poser, Ina

AU - Mann, Matthias

AU - Hyman, Anthony A

AU - Citi, Sandra

PY - 2016/5/20

Y1 - 2016/5/20

N2 - PLEKHA7 is a junctional protein implicated in stabilization of the cadherin protein complex, hypertension, cardiac contractility, glaucoma, microRNA processing, and susceptibility to bacterial toxins. To gain insight into the molecular basis for the functions of PLEKHA7, we looked for new PLEKHA7 interactors. Here, we report the identification of PDZ domain-containing protein 11 (PDZD11) as a new interactor of PLEKHA7 by yeast two-hybrid screening and by mass spectrometry analysis of PLEKHA7 immunoprecipitates. We show that PDZD11 (17 kDa) is expressed in epithelial and endothelial cells, where it forms a complex with PLEKHA7, as determined by co-immunoprecipitation analysis. The N-terminal Trp-Trp (WW) domain of PLEKHA7 interacts directly with the N-terminal 44 amino acids of PDZD11, as shown by GST-pulldown assays. Immunofluorescence analysis shows that PDZD11 is localized at adherens junctions in a PLEKHA7-dependent manner, because its junctional localization is abolished by knock-out of PLEKHA7, and is rescued by re-expression of exogenous PLEKHA7. The junctional recruitment of nectin-1 and nectin-3 and their protein levels are decreased via proteasome-mediated degradation in epithelial cells where either PDZD11 or PLEKHA7 have been knocked-out. PDZD11 forms a complex with nectin-1 and nectin-3, and its PDZ domain interacts directly with the PDZ-binding motif of nectin-1. PDZD11 is required for the efficient assembly of apical junctions of epithelial cells at early time points in the calcium-switch model. These results show that the PLEKHA7-PDZD11 complex stabilizes nectins to promote efficient early junction assembly and uncover a new molecular mechanism through which PLEKHA7 recruits PDZ-binding membrane proteins to epithelial adherens junctions.

AB - PLEKHA7 is a junctional protein implicated in stabilization of the cadherin protein complex, hypertension, cardiac contractility, glaucoma, microRNA processing, and susceptibility to bacterial toxins. To gain insight into the molecular basis for the functions of PLEKHA7, we looked for new PLEKHA7 interactors. Here, we report the identification of PDZ domain-containing protein 11 (PDZD11) as a new interactor of PLEKHA7 by yeast two-hybrid screening and by mass spectrometry analysis of PLEKHA7 immunoprecipitates. We show that PDZD11 (17 kDa) is expressed in epithelial and endothelial cells, where it forms a complex with PLEKHA7, as determined by co-immunoprecipitation analysis. The N-terminal Trp-Trp (WW) domain of PLEKHA7 interacts directly with the N-terminal 44 amino acids of PDZD11, as shown by GST-pulldown assays. Immunofluorescence analysis shows that PDZD11 is localized at adherens junctions in a PLEKHA7-dependent manner, because its junctional localization is abolished by knock-out of PLEKHA7, and is rescued by re-expression of exogenous PLEKHA7. The junctional recruitment of nectin-1 and nectin-3 and their protein levels are decreased via proteasome-mediated degradation in epithelial cells where either PDZD11 or PLEKHA7 have been knocked-out. PDZD11 forms a complex with nectin-1 and nectin-3, and its PDZ domain interacts directly with the PDZ-binding motif of nectin-1. PDZD11 is required for the efficient assembly of apical junctions of epithelial cells at early time points in the calcium-switch model. These results show that the PLEKHA7-PDZD11 complex stabilizes nectins to promote efficient early junction assembly and uncover a new molecular mechanism through which PLEKHA7 recruits PDZ-binding membrane proteins to epithelial adherens junctions.

KW - Adherens Junctions

KW - Animals

KW - Carrier Proteins

KW - Cell Adhesion Molecules

KW - Dogs

KW - Humans

KW - Madin Darby Canine Kidney Cells

KW - Multiprotein Complexes

KW - Nectins

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1074/jbc.m115.712935

DO - 10.1074/jbc.m115.712935

M3 - Journal article

C2 - 27044745

SN - 0021-9258

VL - 291

SP - 11016

EP - 11029

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 21

ER -

PLEKHA7 Recruits PDZD11 to Adherens Junctions to Stabilize Nectins

Abstract

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