Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation

  1. Derek M Britain
  2. Jason P Town
  3. Orion David Weiner  Is a corresponding author
  1. University of California, San Francisco, United States

Abstract

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 (Tischer 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.

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"Figure 4 - Source Data" and "Figure 5 - Source Data" files contain the numerical data used to generate the figures.

Article and author information

Author details

  1. Derek M Britain

    Department of Biochemistry and Biophysics, University of California, San Francisco, San Francsico, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3139-3797
  2. Jason P Town

    Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Orion David Weiner

    Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States
    For correspondence
    orion.weiner@ucsf.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1778-6543

Funding

National Institutes of Health (GM118167)

  • Orion David Weiner

National Science Foundation (DBI-1548297)

  • Orion David Weiner

Novo Nordisk Foundation Center for Basic Metabolic Research (NNF17OC0028176)

  • Orion David Weiner

National Science Foundation (Predoctoral Fellowship)

  • Jason P Town

Achievement Rewards for College Scientists Foundation (Predoctoral Fellowship)

  • Derek M Britain

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Michael L Dustin, University of Oxford, United Kingdom

Publication history

  1. Received: November 10, 2021
  2. Accepted: September 18, 2022
  3. Accepted Manuscript published: September 20, 2022 (version 1)

Copyright

© 2022, Britain et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Derek M Britain
  2. Jason P Town
  3. Orion David Weiner
(2022)
Progressive enhancement of kinetic proofreading in T cell antigen discrimination from receptor activation to DAG generation
eLife 11:e75263.
https://doi.org/10.7554/eLife.75263

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