Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation
T cells use kinetic proofreading to discriminate antigens by converting small changes in antigen-binding lifetime into large differences in cell activation, but where in the signaling cascade this computation is performed is unknown. Previously, we developed a light-gated immune receptor to probe th...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-09-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/75263 |
| _version_ | 1811229369213911040 |
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| author | Derek M Britain Jason P Town Orion David Weiner |
| author_facet | Derek M Britain Jason P Town Orion David Weiner |
| author_sort | Derek M Britain |
| collection | DOAJ |
| description | T cells use kinetic proofreading to discriminate antigens by converting small changes in antigen-binding lifetime into large differences in cell activation, but where in the signaling cascade this computation is performed is unknown. Previously, we developed a light-gated immune receptor to probe the role of ligand kinetics in T cell antigen signaling. We found significant kinetic proofreading at the level of the signaling lipid diacylglycerol (DAG) but lacked the ability to determine where the multiple signaling steps required for kinetic discrimination originate in the upstream signaling cascade (Tiseher and Weiner, 2019). Here, we uncover where kinetic proofreading is executed by adapting our optogenetic system for robust activation of early signaling events. We find the strength of kinetic proofreading progressively increases from Zap70 recruitment to LAT clustering to downstream DAG generation. Leveraging the ability of our system to rapidly disengage ligand binding, we also measure slower reset rates for downstream signaling events. These data suggest a distributed kinetic proofreading mechanism, with proofreading steps both at the receptor and at slower resetting downstream signaling complexes that could help balance antigen sensitivity and discrimination. |
| first_indexed | 2024-04-12T10:13:50Z |
| format | Article |
| id | doaj.art-afa42d8f520241b6ab77a76b12e61144 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T10:13:50Z |
| publishDate | 2022-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-afa42d8f520241b6ab77a76b12e611442022-12-22T03:37:15ZengeLife Sciences Publications LtdeLife2050-084X2022-09-011110.7554/eLife.75263Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generationDerek M Britain0https://orcid.org/0000-0002-3139-3797Jason P Town1Orion David Weiner2https://orcid.org/0000-0002-1778-6543Cardiovascular Research Institute and Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesCardiovascular Research Institute and Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesCardiovascular Research Institute and Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United StatesT cells use kinetic proofreading to discriminate antigens by converting small changes in antigen-binding lifetime into large differences in cell activation, but where in the signaling cascade this computation is performed is unknown. Previously, we developed a light-gated immune receptor to probe the role of ligand kinetics in T cell antigen signaling. We found significant kinetic proofreading at the level of the signaling lipid diacylglycerol (DAG) but lacked the ability to determine where the multiple signaling steps required for kinetic discrimination originate in the upstream signaling cascade (Tiseher and Weiner, 2019). Here, we uncover where kinetic proofreading is executed by adapting our optogenetic system for robust activation of early signaling events. We find the strength of kinetic proofreading progressively increases from Zap70 recruitment to LAT clustering to downstream DAG generation. Leveraging the ability of our system to rapidly disengage ligand binding, we also measure slower reset rates for downstream signaling events. These data suggest a distributed kinetic proofreading mechanism, with proofreading steps both at the receptor and at slower resetting downstream signaling complexes that could help balance antigen sensitivity and discrimination.https://elifesciences.org/articles/75263T cellskinetic proofreadingoptogeneticschimeric antigen receptorantigen discriminationLOV2 |
| spellingShingle | Derek M Britain Jason P Town Orion David Weiner Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation eLife T cells kinetic proofreading optogenetics chimeric antigen receptor antigen discrimination LOV2 |
| title | Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation |
| title_full | Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation |
| title_fullStr | Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation |
| title_full_unstemmed | Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation |
| title_short | Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation |
| title_sort | progressive enhancement of kinetic proofreading in t cell antigen discrimination from receptor activation to dag generation |
| topic | T cells kinetic proofreading optogenetics chimeric antigen receptor antigen discrimination LOV2 |
| url | https://elifesciences.org/articles/75263 |
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