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dc.contributor.authorGallagher, Michael P.
dc.contributor.authorConley, James M.
dc.contributor.authorVangala, Pranitha
dc.contributor.authorReboldi, Andrea
dc.contributor.authorGarber, Manuel
dc.contributor.authorBerg, Leslie J.
dc.date2022-08-11T08:08:25.000
dc.date.accessioned2022-08-23T15:54:48Z
dc.date.available2022-08-23T15:54:48Z
dc.date.issued2020-11-14
dc.date.submitted2020-12-07
dc.identifier.citation<p>bioRxiv 2020.11.12.380725; doi: https://doi.org/10.1101/2020.11.12.380725. <a href="https://doi.org/10.1101/2020.11.12.380725" target="_blank" title="preprint in bioRxiv"> Link to preprint on bioRxiv.</a></p>
dc.identifier.doi10.1101/2020.11.12.380725
dc.identifier.urihttp://hdl.handle.net/20.500.14038/29624
dc.description<p>This article is a preprint. Preprints are preliminary reports of work that have not been certified by peer review.</p>
dc.description.abstractThe strength of peptide:MHC interactions with the T cell receptor (TCR) is correlated with the time to first cell division, the relative scale of the effector cell response, and the graded expression of activation-associated proteins like IRF4. To regulate T cell activation programming, the TCR and the TCR proximal kinase ITK simultaneously trigger many biochemically separate TCR signaling cascades. T cells lacking ITK exhibit selective impairments in effector T cell responses after activation, but under the strongest signaling conditions ITK activity is dispensable. To gain insight into whether TCR signal strength and ITK activity tune observed graded gene expression through unequal activation of disparate signaling pathways, we examined Erk1/2 activation and NFAT, NF-κB translocation in naive OT-I CD8+ cell nuclei. We observed consistent digital activation of NFAT1 and Erk-MAPK, but NF-κB displayed dynamic, graded activation in response to variation in TCR signal strength and was tunable by treatment with an ITK inhibitor. Inhibitor-treated cells showed dampened induction of AP-1 factors Fos and Fosb, NF-κB response gene transcripts, and survival factor Il2 transcripts. ATAC-seq analysis also revealed genomic regions most sensitive to ITK inhibition were enriched for NF-κB and AP-1 motifs. Specific inhibition of NF-κB during peptide stimulation tuned expression of early gene products like c-Fos. Together, these data indicate a key role for ITK in orchestrating optimal activation of separate TCR downstream pathways, specifically aiding NF-κB activation. More broadly, we revealed a mechanism by which variation in TCR signal strength can produce patterns of graded gene expression in activated T cells.
dc.language.isoen_US
dc.relationNow published in Proceedings of the National Academy of Sciences doi: 10.1073/pnas.2025825118
dc.rightsThe copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectImmunology
dc.subjectgene induction
dc.subjectTec kinase ITK
dc.subjectT cell activation
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectCell and Developmental Biology
dc.subjectEnzymes and Coenzymes
dc.subjectImmunopathology
dc.titleThe Tec kinase ITK differentially optimizes NFAT, NF-κB, and MAPK signaling during early T cell activation to regulate graded gene induction [preprint]
dc.typePreprint
dc.source.journaltitlebioRxiv
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=2868&amp;context=faculty_pubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/faculty_pubs/1841
dc.identifier.contextkey20466539
refterms.dateFOA2022-08-23T15:54:48Z
html.description.abstract<p><p id="x-x-x-x-p-2">The strength of peptide:MHC interactions with the T cell receptor (TCR) is correlated with the time to first cell division, the relative scale of the effector cell response, and the graded expression of activation-associated proteins like IRF4. To regulate T cell activation programming, the TCR and the TCR proximal kinase ITK simultaneously trigger many biochemically separate TCR signaling cascades. T cells lacking ITK exhibit selective impairments in effector T cell responses after activation, but under the strongest signaling conditions ITK activity is dispensable. To gain insight into whether TCR signal strength and ITK activity tune observed graded gene expression through unequal activation of disparate signaling pathways, we examined Erk1/2 activation and NFAT, NF-κB translocation in naive OT-I CD8<sup>+</sup> cell nuclei. We observed consistent digital activation of NFAT1 and Erk-MAPK, but NF-κB displayed dynamic, graded activation in response to variation in TCR signal strength and was tunable by treatment with an ITK inhibitor. Inhibitor-treated cells showed dampened induction of AP-1 factors <em>Fos</em> and <em>Fosb</em>, NF-κB response gene transcripts, and survival factor <em>Il2</em> transcripts. ATAC-seq analysis also revealed genomic regions most sensitive to ITK inhibition were enriched for NF-κB and AP-1 motifs. Specific inhibition of NF-κB during peptide stimulation tuned expression of early gene products like c-Fos. Together, these data indicate a key role for ITK in orchestrating optimal activation of separate TCR downstream pathways, specifically aiding NF-κB activation. More broadly, we revealed a mechanism by which variation in TCR signal strength can produce patterns of graded gene expression in activated T cells.</p>
dc.identifier.submissionpathfaculty_pubs/1841
dc.contributor.departmentGarber Lab
dc.contributor.departmentProgram in Bioinformatics and Integrative Biology
dc.contributor.departmentProgram in Molecular Medicine
dc.contributor.departmentDepartment of Pathology
dc.contributor.departmentGraduate School of Biomedical Sciences


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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
Except where otherwise noted, this item's license is described as The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.