Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes
We analyzed here how formin-like 1 β (FMNL1β), an actin cytoskeleton-regulatory protein, regulates microtubule-organizing center (MTOC) and multivesicular bodies (MVB) polarization and exosome secretion at an immune synapse (IS) model in a phosphorylation-dependent manner. IS formation was associate...
| Autores: | , , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2024 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/373082 |
| Acceso en línea: | http://hdl.handle.net/10261/373082 https://api.elsevier.com/content/abstract/scopus_id/85208161032 |
| Access Level: | acceso abierto |
| Palabra clave: | FMNL1β T lymphocytes Actin cytoskeleton Exosomes Human Immune synapse Immunology Inflammation Secretory traffic |
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| dc.title.none.fl_str_mv |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes |
| title |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes |
| spellingShingle |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes Ruiz-Navarro, Javier FMNL1β T lymphocytes Actin cytoskeleton Exosomes Human Immune synapse Immunology Inflammation Secretory traffic |
| title_short |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes |
| title_full |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes |
| title_fullStr |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes |
| title_full_unstemmed |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes |
| title_sort |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytes |
| dc.creator.none.fl_str_mv |
Ruiz-Navarro, Javier Fernández-Hermira, Sara Sanz-Fernández, Irene Barbeito, Pablo Navarro-Zapata, Alfonso Pérez-Martínez, Antonio Garcia-Gonzalo, Francesc R. Calvo, Victor Izquierdo, Manuel |
| author |
Ruiz-Navarro, Javier |
| author_facet |
Ruiz-Navarro, Javier Fernández-Hermira, Sara Sanz-Fernández, Irene Barbeito, Pablo Navarro-Zapata, Alfonso Pérez-Martínez, Antonio Garcia-Gonzalo, Francesc R. Calvo, Victor Izquierdo, Manuel |
| author_role |
author |
| author2 |
Fernández-Hermira, Sara Sanz-Fernández, Irene Barbeito, Pablo Navarro-Zapata, Alfonso Pérez-Martínez, Antonio Garcia-Gonzalo, Francesc R. Calvo, Victor Izquierdo, Manuel |
| author2_role |
author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Ministerio de Ciencia, Innovación y Universidades (España) Agencia Estatal de Investigación (España) Comunidad de Madrid Ruiz-Navarro, Javier [0009-0005-8393-0450] Barbeito, Pablo [0000-0003-0758-0012] Garcia-Gonzalo, Francesc R [0000-0002-9152-2191] Izquierdo Pastor, Manuel [0000-0002-7701-1002] Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72] |
| dc.subject.none.fl_str_mv |
FMNL1β T lymphocytes Actin cytoskeleton Exosomes Human Immune synapse Immunology Inflammation Secretory traffic |
| topic |
FMNL1β T lymphocytes Actin cytoskeleton Exosomes Human Immune synapse Immunology Inflammation Secretory traffic |
| description |
We analyzed here how formin-like 1 β (FMNL1β), an actin cytoskeleton-regulatory protein, regulates microtubule-organizing center (MTOC) and multivesicular bodies (MVB) polarization and exosome secretion at an immune synapse (IS) model in a phosphorylation-dependent manner. IS formation was associated with transient recruitment of FMNL1β to the IS, which was independent of protein kinase C δ (PKCδ). Simultaneous RNA interference of all FMNL1 isoforms prevented MTOC/MVB polarization and exosome secretion, which were restored by FMNL1βWT expression. However, expression of the non-phosphorylatable mutant FMNL1βS1086A did not restore neither MTOC/MVB polarization nor exosome secretion to control levels, supporting the crucial role of S1086 phosphorylation in MTOC/MVB polarization and exosome secretion. In contrast, the phosphomimetic mutant, FMNL1βS1086D, restored MTOC/MVB polarization and exosome secretion. Conversely, FMNL1βS1086D mutant did not recover the deficient MTOC/MVB polarization occurring in PKCδ-interfered clones, indicating that S1086 FMNL1β phosphorylation alone is not sufficient for MTOC/MVB polarization and exosome secretion. FMNL1 interference inhibited the depletion of F-actin at the central region of the immune synapse (cIS), which is necessary for MTOC/MVB polarization. FMNL1βWT and FMNL1βS1086D, but not FMNL1βS1086A expression, restored F-actin depletion at the cIS. Thus, actin cytoskeleton reorganization at the IS underlies the effects of all these FMNL1β variants on polarized secretory traffic. FMNL1 was found in the IS made by primary T lymphocytes, both in T cell receptor (TCR) and chimeric antigen receptor (CAR)-evoked synapses. Taken together, these results point out a crucial role of S1086 phosphorylation in FMNL1β activation, leading to cortical actin reorganization and subsequent control of MTOC/MVB polarization and exosome secretion. |
| publishDate |
2024 |
| dc.date.none.fl_str_mv |
2024 2024 2024 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article http://purl.org/coar/resource_type/c_6501 Publisher's version info:eu-repo/semantics/publishedVersion |
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article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10261/373082 https://api.elsevier.com/content/abstract/scopus_id/85208161032 |
| url |
http://hdl.handle.net/10261/373082 https://api.elsevier.com/content/abstract/scopus_id/85208161032 |
| dc.language.none.fl_str_mv |
Inglés |
| language_invalid_str_mv |
Inglés |
| dc.relation.none.fl_str_mv |
#PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# S2022/BMD-7225 info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-114148RB-I00 info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-104941RB-I00 The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.7554/eLife.96942.4 https://doi.org/10.7554/eLife.96942.4 Sí |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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eLife Sciences Publications |
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eLife Sciences Publications |
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reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC instname:Consejo Superior de Investigaciones Científicas (CSIC) |
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Consejo Superior de Investigaciones Científicas (CSIC) |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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1869425030964707328 |
| spelling |
Formin-like 1β phosphorylation at S1086 is necessary for secretory polarized traffic of exosomes at the immune synapse in Jurkat T lymphocytesRuiz-Navarro, JavierFernández-Hermira, SaraSanz-Fernández, IreneBarbeito, PabloNavarro-Zapata, AlfonsoPérez-Martínez, AntonioGarcia-Gonzalo, Francesc R.Calvo, VictorIzquierdo, ManuelFMNL1βT lymphocytesActin cytoskeletonExosomesHumanImmune synapseImmunologyInflammationSecretory trafficWe analyzed here how formin-like 1 β (FMNL1β), an actin cytoskeleton-regulatory protein, regulates microtubule-organizing center (MTOC) and multivesicular bodies (MVB) polarization and exosome secretion at an immune synapse (IS) model in a phosphorylation-dependent manner. IS formation was associated with transient recruitment of FMNL1β to the IS, which was independent of protein kinase C δ (PKCδ). Simultaneous RNA interference of all FMNL1 isoforms prevented MTOC/MVB polarization and exosome secretion, which were restored by FMNL1βWT expression. However, expression of the non-phosphorylatable mutant FMNL1βS1086A did not restore neither MTOC/MVB polarization nor exosome secretion to control levels, supporting the crucial role of S1086 phosphorylation in MTOC/MVB polarization and exosome secretion. In contrast, the phosphomimetic mutant, FMNL1βS1086D, restored MTOC/MVB polarization and exosome secretion. Conversely, FMNL1βS1086D mutant did not recover the deficient MTOC/MVB polarization occurring in PKCδ-interfered clones, indicating that S1086 FMNL1β phosphorylation alone is not sufficient for MTOC/MVB polarization and exosome secretion. FMNL1 interference inhibited the depletion of F-actin at the central region of the immune synapse (cIS), which is necessary for MTOC/MVB polarization. FMNL1βWT and FMNL1βS1086D, but not FMNL1βS1086A expression, restored F-actin depletion at the cIS. Thus, actin cytoskeleton reorganization at the IS underlies the effects of all these FMNL1β variants on polarized secretory traffic. FMNL1 was found in the IS made by primary T lymphocytes, both in T cell receptor (TCR) and chimeric antigen receptor (CAR)-evoked synapses. Taken together, these results point out a crucial role of S1086 phosphorylation in FMNL1β activation, leading to cortical actin reorganization and subsequent control of MTOC/MVB polarization and exosome secretion.Work in the F.R.G-G lab was funded by grant PID2019-104941RB-I00 from the Spanish Ministry of Science and Innovation (PID2019-104941RB-I00/AEI/10.13039/501100011033). This work was supported by a grant from the Programa Estatal de Investigación, Desarrollo e Innovación, Modalidad Retos Investigación (Grant PID2020-114148RB-I00) funded by Spanish Ministry of Science and Innovation (PID2020-114148RB-I00/AEI/10.13039/501100011033), and grant P2022/BMD-7225, funded by Consortia in Biomedicine of Comunidad de Madrid to MI.Peer reviewedeLife Sciences PublicationsMinisterio de Ciencia, Innovación y Universidades (España)Agencia Estatal de Investigación (España)Comunidad de MadridRuiz-Navarro, Javier [0009-0005-8393-0450]Barbeito, Pablo [0000-0003-0758-0012]Garcia-Gonzalo, Francesc R [0000-0002-9152-2191]Izquierdo Pastor, Manuel [0000-0002-7701-1002]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242024info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/373082https://api.elsevier.com/content/abstract/scopus_id/85208161032reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#S2022/BMD-7225info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-114148RB-I00info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-104941RB-I00The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.7554/eLife.96942.4https://doi.org/10.7554/eLife.96942.4Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3730822026-05-22T06:33:51Z |
| score |
15,811543 |