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Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse

Diacylglycerol (DAG) generation at the T cell immunological synapse (IS) determines the correct activation of antigen-specific immune responses. DAG kinases (DGKs) α and ζ act as negative regulators of DAG-mediated signals by catalyzing DAG conversion to phosphatidic acid (PA). Nonetheless, the spec...

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Autores principales: Gharbi, Severine I., Rincón, Esther, Avila-Flores, Antonia, Torres-Ayuso, Pedro, Almena, María, Cobos, María Angeles, Albar, Juan Pablo, Mérida, Isabel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3216665/
https://www.ncbi.nlm.nih.gov/pubmed/21937721
http://dx.doi.org/10.1091/mbc.E11-03-0247
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author Gharbi, Severine I.
Rincón, Esther
Avila-Flores, Antonia
Torres-Ayuso, Pedro
Almena, María
Cobos, María Angeles
Albar, Juan Pablo
Mérida, Isabel
author_facet Gharbi, Severine I.
Rincón, Esther
Avila-Flores, Antonia
Torres-Ayuso, Pedro
Almena, María
Cobos, María Angeles
Albar, Juan Pablo
Mérida, Isabel
author_sort Gharbi, Severine I.
collection PubMed
description Diacylglycerol (DAG) generation at the T cell immunological synapse (IS) determines the correct activation of antigen-specific immune responses. DAG kinases (DGKs) α and ζ act as negative regulators of DAG-mediated signals by catalyzing DAG conversion to phosphatidic acid (PA). Nonetheless, the specific input of each enzyme and their spatial regulation during IS formation remain uncharacterized. Here we report recruitment of endogenous DGKα and DGKζ to the T cell receptor (TCR) complex following TCR/CD28 engagement. Specific DGK gene silencing shows that PA production at the activated complex depends mainly on DGKζ, indicating functional differences between these proteins. DGKζ kinase activity at the TCR is enhanced by phorbol-12-myristate-13-acetate cotreatment, suggesting DAG-mediated regulation of DGKζ responsiveness. We used GFP-DGKζ and -DGKα chimeras to assess translocation dynamics during IS formation. Only GFP-DGKζ translocated rapidly to the plasma membrane at early stages of IS formation, independent of enzyme activity. Finally, use of a fluorescent DAG sensor confirmed rapid, sustained DAG accumulation at the IS and allowed us to directly correlate membrane translocation of active DGKζ with DAG consumption at the IS. This study highlights a DGKζ-specific function for local DAG metabolism at the IS and offers new clues to its mode of regulation.
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spelling pubmed-32166652012-01-30 Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse Gharbi, Severine I. Rincón, Esther Avila-Flores, Antonia Torres-Ayuso, Pedro Almena, María Cobos, María Angeles Albar, Juan Pablo Mérida, Isabel Mol Biol Cell Articles Diacylglycerol (DAG) generation at the T cell immunological synapse (IS) determines the correct activation of antigen-specific immune responses. DAG kinases (DGKs) α and ζ act as negative regulators of DAG-mediated signals by catalyzing DAG conversion to phosphatidic acid (PA). Nonetheless, the specific input of each enzyme and their spatial regulation during IS formation remain uncharacterized. Here we report recruitment of endogenous DGKα and DGKζ to the T cell receptor (TCR) complex following TCR/CD28 engagement. Specific DGK gene silencing shows that PA production at the activated complex depends mainly on DGKζ, indicating functional differences between these proteins. DGKζ kinase activity at the TCR is enhanced by phorbol-12-myristate-13-acetate cotreatment, suggesting DAG-mediated regulation of DGKζ responsiveness. We used GFP-DGKζ and -DGKα chimeras to assess translocation dynamics during IS formation. Only GFP-DGKζ translocated rapidly to the plasma membrane at early stages of IS formation, independent of enzyme activity. Finally, use of a fluorescent DAG sensor confirmed rapid, sustained DAG accumulation at the IS and allowed us to directly correlate membrane translocation of active DGKζ with DAG consumption at the IS. This study highlights a DGKζ-specific function for local DAG metabolism at the IS and offers new clues to its mode of regulation. The American Society for Cell Biology 2011-11-15 /pmc/articles/PMC3216665/ /pubmed/21937721 http://dx.doi.org/10.1091/mbc.E11-03-0247 Text en © 2011 Gharbi et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
Gharbi, Severine I.
Rincón, Esther
Avila-Flores, Antonia
Torres-Ayuso, Pedro
Almena, María
Cobos, María Angeles
Albar, Juan Pablo
Mérida, Isabel
Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse
title Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse
title_full Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse
title_fullStr Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse
title_full_unstemmed Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse
title_short Diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse
title_sort diacylglycerol kinase ζ controls diacylglycerol metabolism at the immunological synapse
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3216665/
https://www.ncbi.nlm.nih.gov/pubmed/21937721
http://dx.doi.org/10.1091/mbc.E11-03-0247
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