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dc.contributor.authorJahan, Akhee S.
dc.contributor.authorLestra, Maxime
dc.contributor.authorSwee, Lee Kim
dc.contributor.authorFan, Ying
dc.contributor.authorLamers, Mart M.
dc.contributor.authorTafesse, Fikadu G.
dc.contributor.authorTheile, Christopher S.
dc.contributor.authorSpooner, Eric
dc.contributor.authorBruzzone, Roberto
dc.contributor.authorSanyal, Sumana
dc.contributor.authorPloegh, Hidde
dc.date.accessioned2017-01-12T20:56:28Z
dc.date.available2017-01-12T20:56:28Z
dc.date.issued2016-02
dc.date.submitted2015-11
dc.identifier.issn0027-8424
dc.identifier.issn1091-6490
dc.identifier.urihttp://hdl.handle.net/1721.1/106472
dc.description.abstractPosttranslational modifications are central to the spatial and temporal regulation of protein function. Among others, phosphorylation and ubiquitylation are known to regulate proximal T-cell receptor (TCR) signaling. Here we used a systematic and unbiased approach to uncover deubiquitylating enzymes (DUBs) that participate during TCR signaling in primary mouse T lymphocytes. Using a C-terminally modified vinyl methyl ester variant of ubiquitin (HA-Ub-VME), we captured DUBs that are differentially recruited to the cytosol on TCR activation. We identified ubiquitin-specific peptidase (Usp) 12 and Usp46, which had not been previously described in this pathway. Stimulation with anti-CD3 resulted in phosphorylation and time-dependent translocation of Usp12 from the nucleus to the cytosol. Usp12−/− Jurkat cells displayed defective NFκB, NFAT, and MAPK activities owing to attenuated surface expression of TCR, which were rescued on reconstitution of wild type Usp12. Proximity-based labeling with BirA-Usp12 revealed several TCR adaptor proteins acting as interactors in stimulated cells, of which LAT and Trat1 displayed reduced expression in Usp12−/− cells. We demonstrate that Usp12 deubiquitylates and prevents lysosomal degradation of LAT and Trat1 to maintain the proximal TCR complex for the duration of signaling. Our approach benefits from the use of activity-based probes in primary cells without any previous genome modification, and underscores the importance of ubiquitin-mediated regulation to refine signaling cascades.en_US
dc.description.sponsorshipInstitut Pasteur International Networken_US
dc.description.sponsorshipUniversity Grants Committee (Hong Kong, China) (Grant AoE/M-12/05)en_US
dc.language.isoen_US
dc.publisherNational Academy of Sciences (U.S.)en_US
dc.relation.isversionofhttp://dx.doi.org/10.1073/pnas.1521763113en_US
dc.rightsArticle is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.en_US
dc.sourcePNASen_US
dc.titleUsp12 stabilizes the T-cell receptor complex at the cell surface during signalingen_US
dc.typeArticleen_US
dc.identifier.citationJahan, Akhee S. et al. “Usp12 Stabilizes the T-Cell Receptor Complex at the Cell Surface during Signaling.” Proceedings of the National Academy of Sciences 113.6 (2016): E705–E714. © 2016 National Academy of Sciencesen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biologyen_US
dc.contributor.departmentWhitehead Institute for Biomedical Researchen_US
dc.contributor.mitauthorPloegh, Hidde
dc.relation.journalProceedings of the National Academy of Sciencesen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsJahan, Akhee S.; Lestra, Maxime; Swee, Lee Kim; Fan, Ying; Lamers, Mart M.; Tafesse, Fikadu G.; Theile, Christopher S.; Spooner, Eric; Bruzzone, Roberto; Ploegh, Hidde L.; Sanyal, Sumanaen_US
dspace.embargo.termsNen_US
dc.identifier.orcidhttps://orcid.org/0000-0002-1090-6071
mit.licensePUBLISHER_POLICYen_US


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