TRPC6 binds to and activates calpain, independent of its channel activity, and regulates podocyte cytoskeleton, cell adhesion, and motility

Background: Mutations in transient receptor potential channel 6 (TRPC6) are associated with an inherited form of focal segmental glomerulosclerosis (FSGS). Despite widespread expression, patients with TRPC6 mutations do not present with any other pathological phenotype suggesting that this protein h...

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Autores principales: Farmer, Louise K., Rollason, Ruth, Whitcomb, Daniel J., Ni, Lan, Goodliff, Alexander, Lay, Abigail C., Birnbaumer, Lutz, Heesom, Kate J., Xu, Shang-Zhong, Saleem, Moin A., Welsh, Gavin I.
Otros Autores: 0000-0002-2148-6658
Formato: Artículo
Lenguaje:Inglés
Publicado: American Society of Nephrology 2019
Materias:
NEFROLOGIA
MUTACION
GENES
TRASTORNOS GENETICOS
ENFERMEDADES RENALES
RIÑON
PROTEINAS
Acceso en línea:https://repositorio.uca.edu.ar/handle/123456789/8678
Aporte de:
Repositorio Institucional de la Universidad Católica Argentina (UCA) de Universidad Católica Argentina
id I33-R139123456789-8678
record_format dspace
institution Universidad Católica Argentina
institution_str I-33
repository_str R-139
collection Repositorio Institucional de la Universidad Católica Argentina (UCA)
language Inglés
topic NEFROLOGIA
MUTACION
GENES
TRASTORNOS GENETICOS
ENFERMEDADES RENALES
RIÑON
PROTEINAS
spellingShingle NEFROLOGIA
MUTACION
GENES
TRASTORNOS GENETICOS
ENFERMEDADES RENALES
RIÑON
PROTEINAS
Farmer, Louise K.
Rollason, Ruth
Whitcomb, Daniel J.
Ni, Lan
Goodliff, Alexander
Lay, Abigail C.
Birnbaumer, Lutz
Heesom, Kate J.
Xu, Shang-Zhong
Saleem, Moin A.
Welsh, Gavin I.
TRPC6 binds to and activates calpain, independent of its channel activity, and regulates podocyte cytoskeleton, cell adhesion, and motility
topic_facet NEFROLOGIA
MUTACION
GENES
TRASTORNOS GENETICOS
ENFERMEDADES RENALES
RIÑON
PROTEINAS
description Background: Mutations in transient receptor potential channel 6 (TRPC6) are associated with an inherited form of focal segmental glomerulosclerosis (FSGS). Despite widespread expression, patients with TRPC6 mutations do not present with any other pathological phenotype suggesting that this protein has a unique but yet unidentified role within the target cell for FSGS, the kidney podocyte. Methods: A stable TRPC6 knock out podocyte cell line was generated from TRPC6 knockout mice. These cells were engineered to express wild type, dominant negative or either G109S or K874* disease causing mutants of TRPC6. These cells were extensively characterised via motility, detachment and calpain activity assays, immunofluorescence and confocal or Total Internal Reflection fluorescence (TIRF) microscopy, and western blotting. Results: TRPC6-/- podocytes are less motile and more adhesive, with an altered actin cytoskeleton compared to wild type cells. Mechanistically, we show that TRPC6 binds to ERK1/2 and the actin regulatory proteins, caldesmon and calpain 1 and 2. Calpains are calcium dependent cysteine proteases, which control the podocyte cytoskeleton, cell adhesion and motility via cleavage of paxillin, focal adhesion kinase and talin. Knockdown or expression of the truncated K874*, but not the gain of function G019S or dominant negative mutant of TRPC6 results in the mislocalization of calpain 1 and 2 and significant down-regulation of calpain activity leading to altered podocyte cytoskeleton, motility and adhesion, a phenocopy of TRPC6 -/- podocytes. Conclusions: Our data demonstrates that the physical interaction between TRPC6 and calpain in the podocyte is important in disease, independent of TRPC6 channel activity.
author2 0000-0002-2148-6658
author_facet 0000-0002-2148-6658
Farmer, Louise K.
Rollason, Ruth
Whitcomb, Daniel J.
Ni, Lan
Goodliff, Alexander
Lay, Abigail C.
Birnbaumer, Lutz
Heesom, Kate J.
Xu, Shang-Zhong
Saleem, Moin A.
Welsh, Gavin I.
format Artículo
author Farmer, Louise K.
Rollason, Ruth
Whitcomb, Daniel J.
Ni, Lan
Goodliff, Alexander
Lay, Abigail C.
Birnbaumer, Lutz
Heesom, Kate J.
Xu, Shang-Zhong
Saleem, Moin A.
Welsh, Gavin I.
author_sort Farmer, Louise K.
title TRPC6 binds to and activates calpain, independent of its channel activity, and regulates podocyte cytoskeleton, cell adhesion, and motility
title_short TRPC6 binds to and activates calpain, independent of its channel activity, and regulates podocyte cytoskeleton, cell adhesion, and motility
title_full TRPC6 binds to and activates calpain, independent of its channel activity, and regulates podocyte cytoskeleton, cell adhesion, and motility
title_fullStr TRPC6 binds to and activates calpain, independent of its channel activity, and regulates podocyte cytoskeleton, cell adhesion, and motility
title_full_unstemmed TRPC6 binds to and activates calpain, independent of its channel activity, and regulates podocyte cytoskeleton, cell adhesion, and motility
title_sort trpc6 binds to and activates calpain, independent of its channel activity, and regulates podocyte cytoskeleton, cell adhesion, and motility
publisher American Society of Nephrology
publishDate 2019
url https://repositorio.uca.edu.ar/handle/123456789/8678
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