Cargando…

Recruitment of Slp-76 to the Membrane and Glycolipid-Enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling

Two hematopoietic-specific adapters, src homology 2 domain–containing leukocyte phosphoprotein of 76 kD (SLP-76) and linker for activation of T cells (LAT), are critical for T cell development and T cell receptor (TCR) signaling. Several studies have suggested that SLP-76 and LAT function coordinate...

Descripción completa

Detalles Bibliográficos
Autores principales: Boerth, Nancy J., Sadler, Jeffrey J., Bauer, Daniel E., Clements, James L., Gheith, Shereen M., Koretzky, Gary A.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2000
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2193307/
https://www.ncbi.nlm.nih.gov/pubmed/11015445
_version_ 1782147439986212864
author Boerth, Nancy J.
Sadler, Jeffrey J.
Bauer, Daniel E.
Clements, James L.
Gheith, Shereen M.
Koretzky, Gary A.
author_facet Boerth, Nancy J.
Sadler, Jeffrey J.
Bauer, Daniel E.
Clements, James L.
Gheith, Shereen M.
Koretzky, Gary A.
author_sort Boerth, Nancy J.
collection PubMed
description Two hematopoietic-specific adapters, src homology 2 domain–containing leukocyte phosphoprotein of 76 kD (SLP-76) and linker for activation of T cells (LAT), are critical for T cell development and T cell receptor (TCR) signaling. Several studies have suggested that SLP-76 and LAT function coordinately to promote downstream signaling. In support of this hypothesis, we find that a fraction of SLP-76 localizes to glycolipid-enriched membrane microdomains (GEMs) after TCR stimulation. This recruitment of SLP-76 requires amino acids 224–244. The functional consequences of targeting SLP-76 to GEMs for TCR signaling are demonstrated using a LAT/SLP-76 chimeric protein. Expression of this construct reconstitutes TCR-inducted phospholipase Cγ1 phosphorylation, extracellular signal–regulated kinase activation, and nuclear factor of activated T cells (NFAT) promoter activity in LAT-deficient Jurkat T cells (J.CaM2). Mutation of the chimeric construct precluding its recruitment to GEMs diminishes but does not eliminate its ability to support TCR signaling. Expression of a chimera that lacks SLP-76 amino acids 224–244 restores NFAT promoter activity, suggesting that if localized, SLP-76 does not require an association with Gads to promote T cell activation. In contrast, mutation of the protein tyrosine kinase phosphorylation sites of SLP-76 in the context of the LAT/SLP-76 chimera abolishes reconstitution of TCR function. Collectively, these experiments show that optimal TCR signaling relies on the compartmentalization of SLP-76 and that one critical function of LAT is to bring SLP-76 and its associated proteins to the membrane.
format Text
id pubmed-2193307
institution National Center for Biotechnology Information
language English
publishDate 2000
publisher The Rockefeller University Press
record_format MEDLINE/PubMed
spelling pubmed-21933072008-04-16 Recruitment of Slp-76 to the Membrane and Glycolipid-Enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling Boerth, Nancy J. Sadler, Jeffrey J. Bauer, Daniel E. Clements, James L. Gheith, Shereen M. Koretzky, Gary A. J Exp Med Original Article Two hematopoietic-specific adapters, src homology 2 domain–containing leukocyte phosphoprotein of 76 kD (SLP-76) and linker for activation of T cells (LAT), are critical for T cell development and T cell receptor (TCR) signaling. Several studies have suggested that SLP-76 and LAT function coordinately to promote downstream signaling. In support of this hypothesis, we find that a fraction of SLP-76 localizes to glycolipid-enriched membrane microdomains (GEMs) after TCR stimulation. This recruitment of SLP-76 requires amino acids 224–244. The functional consequences of targeting SLP-76 to GEMs for TCR signaling are demonstrated using a LAT/SLP-76 chimeric protein. Expression of this construct reconstitutes TCR-inducted phospholipase Cγ1 phosphorylation, extracellular signal–regulated kinase activation, and nuclear factor of activated T cells (NFAT) promoter activity in LAT-deficient Jurkat T cells (J.CaM2). Mutation of the chimeric construct precluding its recruitment to GEMs diminishes but does not eliminate its ability to support TCR signaling. Expression of a chimera that lacks SLP-76 amino acids 224–244 restores NFAT promoter activity, suggesting that if localized, SLP-76 does not require an association with Gads to promote T cell activation. In contrast, mutation of the protein tyrosine kinase phosphorylation sites of SLP-76 in the context of the LAT/SLP-76 chimera abolishes reconstitution of TCR function. Collectively, these experiments show that optimal TCR signaling relies on the compartmentalization of SLP-76 and that one critical function of LAT is to bring SLP-76 and its associated proteins to the membrane. The Rockefeller University Press 2000-10-02 /pmc/articles/PMC2193307/ /pubmed/11015445 Text en © 2000 The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Original Article
Boerth, Nancy J.
Sadler, Jeffrey J.
Bauer, Daniel E.
Clements, James L.
Gheith, Shereen M.
Koretzky, Gary A.
Recruitment of Slp-76 to the Membrane and Glycolipid-Enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling
title Recruitment of Slp-76 to the Membrane and Glycolipid-Enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling
title_full Recruitment of Slp-76 to the Membrane and Glycolipid-Enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling
title_fullStr Recruitment of Slp-76 to the Membrane and Glycolipid-Enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling
title_full_unstemmed Recruitment of Slp-76 to the Membrane and Glycolipid-Enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling
title_short Recruitment of Slp-76 to the Membrane and Glycolipid-Enriched Membrane Microdomains Replaces the Requirement for Linker for Activation of T Cells in T Cell Receptor Signaling
title_sort recruitment of slp-76 to the membrane and glycolipid-enriched membrane microdomains replaces the requirement for linker for activation of t cells in t cell receptor signaling
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2193307/
https://www.ncbi.nlm.nih.gov/pubmed/11015445
work_keys_str_mv AT boerthnancyj recruitmentofslp76tothemembraneandglycolipidenrichedmembranemicrodomainsreplacestherequirementforlinkerforactivationoftcellsintcellreceptorsignaling
AT sadlerjeffreyj recruitmentofslp76tothemembraneandglycolipidenrichedmembranemicrodomainsreplacestherequirementforlinkerforactivationoftcellsintcellreceptorsignaling
AT bauerdaniele recruitmentofslp76tothemembraneandglycolipidenrichedmembranemicrodomainsreplacestherequirementforlinkerforactivationoftcellsintcellreceptorsignaling
AT clementsjamesl recruitmentofslp76tothemembraneandglycolipidenrichedmembranemicrodomainsreplacestherequirementforlinkerforactivationoftcellsintcellreceptorsignaling
AT gheithshereenm recruitmentofslp76tothemembraneandglycolipidenrichedmembranemicrodomainsreplacestherequirementforlinkerforactivationoftcellsintcellreceptorsignaling
AT koretzkygarya recruitmentofslp76tothemembraneandglycolipidenrichedmembranemicrodomainsreplacestherequirementforlinkerforactivationoftcellsintcellreceptorsignaling