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eNOS S-nitrosylates β-actin on Cys374 and regulates PKC-θ at the immune synapse by impairing actin binding to profilin-1
The actin cytoskeleton coordinates the organization of signaling microclusters at the immune synapse (IS); however, the mechanisms involved remain poorly understood. We show here that nitric oxide (NO) generated by endothelial nitric oxide synthase (eNOS) controls the coalescence of protein kinase C...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5386235/ https://www.ncbi.nlm.nih.gov/pubmed/28394935 http://dx.doi.org/10.1371/journal.pbio.2000653 |
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author | García-Ortiz, Almudena Martín-Cofreces, Noa B. Ibiza, Sales Ortega, Ángel Izquierdo-Álvarez, Alicia Trullo, Antonio Victor, Víctor M. Calvo, Enrique Sot, Begoña Martínez-Ruiz, Antonio Vázquez, Jesús Sánchez-Madrid, Francisco Serrador, Juan M. |
author_facet | García-Ortiz, Almudena Martín-Cofreces, Noa B. Ibiza, Sales Ortega, Ángel Izquierdo-Álvarez, Alicia Trullo, Antonio Victor, Víctor M. Calvo, Enrique Sot, Begoña Martínez-Ruiz, Antonio Vázquez, Jesús Sánchez-Madrid, Francisco Serrador, Juan M. |
author_sort | García-Ortiz, Almudena |
collection | PubMed |
description | The actin cytoskeleton coordinates the organization of signaling microclusters at the immune synapse (IS); however, the mechanisms involved remain poorly understood. We show here that nitric oxide (NO) generated by endothelial nitric oxide synthase (eNOS) controls the coalescence of protein kinase C-θ (PKC-θ) at the central supramolecular activation cluster (c-SMAC) of the IS. eNOS translocated with the Golgi to the IS and partially colocalized with F-actin around the c-SMAC. This resulted in reduced actin polymerization and centripetal retrograde flow of β-actin and PKC-θ from the lamellipodium-like distal (d)-SMAC, promoting PKC-θ activation. Furthermore, eNOS-derived NO S-nitrosylated β-actin on Cys374 and impaired actin binding to profilin-1 (PFN1), as confirmed with the transnitrosylating agent S-nitroso-L-cysteine (Cys-NO). The importance of NO and the formation of PFN1-actin complexes on the regulation of PKC-θ was corroborated by overexpression of PFN1- and actin-binding defective mutants of β-actin (C374S) and PFN1 (H119E), respectively, which reduced the coalescence of PKC-θ at the c-SMAC. These findings unveil a novel NO-dependent mechanism by which the actin cytoskeleton controls the organization and activation of signaling microclusters at the IS. |
format | Online Article Text |
id | pubmed-5386235 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-53862352017-05-03 eNOS S-nitrosylates β-actin on Cys374 and regulates PKC-θ at the immune synapse by impairing actin binding to profilin-1 García-Ortiz, Almudena Martín-Cofreces, Noa B. Ibiza, Sales Ortega, Ángel Izquierdo-Álvarez, Alicia Trullo, Antonio Victor, Víctor M. Calvo, Enrique Sot, Begoña Martínez-Ruiz, Antonio Vázquez, Jesús Sánchez-Madrid, Francisco Serrador, Juan M. PLoS Biol Research Article The actin cytoskeleton coordinates the organization of signaling microclusters at the immune synapse (IS); however, the mechanisms involved remain poorly understood. We show here that nitric oxide (NO) generated by endothelial nitric oxide synthase (eNOS) controls the coalescence of protein kinase C-θ (PKC-θ) at the central supramolecular activation cluster (c-SMAC) of the IS. eNOS translocated with the Golgi to the IS and partially colocalized with F-actin around the c-SMAC. This resulted in reduced actin polymerization and centripetal retrograde flow of β-actin and PKC-θ from the lamellipodium-like distal (d)-SMAC, promoting PKC-θ activation. Furthermore, eNOS-derived NO S-nitrosylated β-actin on Cys374 and impaired actin binding to profilin-1 (PFN1), as confirmed with the transnitrosylating agent S-nitroso-L-cysteine (Cys-NO). The importance of NO and the formation of PFN1-actin complexes on the regulation of PKC-θ was corroborated by overexpression of PFN1- and actin-binding defective mutants of β-actin (C374S) and PFN1 (H119E), respectively, which reduced the coalescence of PKC-θ at the c-SMAC. These findings unveil a novel NO-dependent mechanism by which the actin cytoskeleton controls the organization and activation of signaling microclusters at the IS. Public Library of Science 2017-04-10 /pmc/articles/PMC5386235/ /pubmed/28394935 http://dx.doi.org/10.1371/journal.pbio.2000653 Text en © 2017 García-Ortiz et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article García-Ortiz, Almudena Martín-Cofreces, Noa B. Ibiza, Sales Ortega, Ángel Izquierdo-Álvarez, Alicia Trullo, Antonio Victor, Víctor M. Calvo, Enrique Sot, Begoña Martínez-Ruiz, Antonio Vázquez, Jesús Sánchez-Madrid, Francisco Serrador, Juan M. eNOS S-nitrosylates β-actin on Cys374 and regulates PKC-θ at the immune synapse by impairing actin binding to profilin-1 |
title | eNOS S-nitrosylates β-actin on Cys374 and regulates PKC-θ at the immune synapse by impairing actin binding to profilin-1 |
title_full | eNOS S-nitrosylates β-actin on Cys374 and regulates PKC-θ at the immune synapse by impairing actin binding to profilin-1 |
title_fullStr | eNOS S-nitrosylates β-actin on Cys374 and regulates PKC-θ at the immune synapse by impairing actin binding to profilin-1 |
title_full_unstemmed | eNOS S-nitrosylates β-actin on Cys374 and regulates PKC-θ at the immune synapse by impairing actin binding to profilin-1 |
title_short | eNOS S-nitrosylates β-actin on Cys374 and regulates PKC-θ at the immune synapse by impairing actin binding to profilin-1 |
title_sort | enos s-nitrosylates β-actin on cys374 and regulates pkc-θ at the immune synapse by impairing actin binding to profilin-1 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5386235/ https://www.ncbi.nlm.nih.gov/pubmed/28394935 http://dx.doi.org/10.1371/journal.pbio.2000653 |
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