Phosphoproteomic Analyses of Interleukin 2 Signaling Reveal Integrated JAK Kinase-Dependent and -Independent Networks in CD8 + T Cells
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Interleukin-2 (IL-2) is a fundamental cytokine that controls proliferation and differentiation of T cells. Here, we used high-resolution mass spectrometry to generate a comprehensive and detailed map of IL-2 protein phosphorylations in cytotoxic T cells (CTL). The data revealed that Janus kinases (JAKs) couple IL-2 receptors to the coordinated phosphorylation of transcription factors, regulators of chromatin, mRNA translation, GTPases, vesicle trafficking, and the actin and microtubule cytoskeleton. We identified an IL-2-JAK-independent SRC family Tyr-kinase-controlled signaling network that regulates ∼10% of the CTL phosphoproteome, the production of phosphatidylinositol (3,4,5)-trisphosphate (PIP3), and the activity of the serine/threonine kinase AKT. These data reveal a signaling framework wherein IL-2-JAK-controlled pathways coordinate with IL-2-independent networks of kinase activity and provide a resource toward the further understanding of the networks of protein phosphorylation that program CTL fate.
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TCR+IL-2 treatment of resting and activated human CD4+ and CD8+ T cells activated a vast array of signaling intermediaries detected by the antibody array, serving as a positive control.
“In both resting and activated human CD4+ and CD8+ T cells, TCR+IL-2 treatment led to the activation of a vast array of signaling intermediaries, including STAT1, STAT3, STAT4, STAT5, STAT6, ERK, AKT, S6R, CREB”
From CDK8 fine-tunes IL-6 transcriptional activities by limiting STAT3 resident time at the gene loci (2020) · cites this paper as Ross et al., 2016
The Results section (‘The IL-2-Regulated Phosphoproteome’; page unavailable) reports that IL-2 stimulation changed many phosphorylations in CD8+ cytotoxic T cells, including STAT5 and ERK. It does not establish the claimed response to combined TCR+IL-2 treatment in resting and activated human CD4+ and CD8+ cells; the study used mass spectrometry rather than an antibody array and does not support the full list of intermediaries. (LLM, conf=0.88)
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Finlay, D. K., Rosenzweig, E., Sinclair, L. V., Feijoo-Carnero, C., Hukelmann, J. L., Rolf, J., Panteleyev, A. A., Okkenhaug, K., & Cantrell, D. A. (2012) · Journal of Experimental Medicine · doi:10.1084/jem.20112607 · Cited by 596 · see citing works
Self-citationLu, W., Gong, D., Bar-Sagi, D., & Cole, P. A. (2001) · Molecular Cell · doi:10.1016/s1097-2765(01)00369-0 · Cited by 268 · see citing works
The cytotoxic T cell proteome and its shaping by the kinase mTOR
Hukelmann, J. L., Anderson, K. E., Sinclair, L. V., Grzes, K. M., Murillo, A. B., Hawkins, P. T., Stephens, L. R., Lamond, A. I., & Cantrell, D. A. (2016) · Nature Immunology · doi:10.1038/ni.3314 · Cited by 249 · see citing works
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Research works printed but not linked: 54
- Angulo I. Vadas O. Garçon F. Banham-Hall E. Plagnol V. Leahy T.R. Baxendale H. Coulter T. Curtis J. Wu C. Phosphoinositide 3-kinase δ gene mutation predisposes to respiratory infection and airway damage Science 342 2013 866 871 24136356 PMC3930011
- Arenas-Ramirez N. Woytschak J. Boyman O. Interleukin-2: biology, design and application Trends Immunol. 36 2015 763 777 26572555
- Arneja A. Johnson H. Gabrovsek L. Lauffenburger D.A. White F.M. Qualitatively different T cell phenotypic responses to IL-2 versus IL-15 are unified by identical dependences on receptor signal strength and duration J. Immunol. 192 2014 123 135 24298013 PMC3950894
- Beadling C. Ng J. Babbage J.W. Cantrell D.A. Interleukin-2 activation of STAT5 requires the convergent action of tyrosine kinases and a serine/threonine kinase pathway distinct from the Raf1/ERK2 MAP kinase pathway EMBO J. 15 1996 1902 1913 8617237 PMC450109
- Bensinger S.J. Walsh P.T. Zhang J. Carroll M. Parsons R. Rathmell J.C. Thompson C.B. Burchill M.A. Farrar M.A. Turka L.A. Distinct IL-2 receptor signaling pattern in CD4+CD25+ regulatory T cells J. Immunol. 172 2004 5287 5296 15100267 PMC2842445
- Chamorro M. Czar M.J. Debnath J. Cheng G. Lenardo M.J. Varmus H.E. Schwartzberg P.L. Requirements for activation and RAFT localization of the T-lymphocyte kinase Rlk/Txk BMC Immunol. 2 2001 3 11353545 PMC31577
- Chang C.-W. Chou H.-Y. Lin Y.-S. Huang K.-H. Chang C.-J. Hsu T.-C. Lee S.-C. Phosphorylation at Ser473 regulates heterochromatin protein 1 binding and corepressor function of TIF1beta/KAP1 BMC Mol. Biol. 9 2008 61 18590578 PMC2474647
- Chang V.T. Fernandes R.A. Ganzinger K.A. Lee S.F. Siebold C. McColl J. Jönsson P. Palayret M. Harlos K. Coles C.H. Initiation of T cell signaling by CD45 segregation at ‘close contacts’ Nat. Immunol. 17 2016 574 582 26998761 PMC4839504
- Chikuma S. Suita N. Okazaki I.-M. Shibayama S. Honjo T. TRIM28 prevents autoinflammatory T cell development in vivo Nat. Immunol. 13 2012 596 603 22544392
- Clark D.E. Williams C.C. Duplessis T.T. Moring K.L. Notwick A.R. Long W. Lane W.S. Beuvink I. Hynes N.E. Jones F.E. ERBB4/HER4 potentiates STAT5A transcriptional activity by regulating novel STAT5A serine phosphorylation events J. Biol. Chem. 280 2005 24175 24180 15863494
- Cornish G.H. Sinclair L.V. Cantrell D.A. Differential regulation of T-cell growth by IL-2 and IL-15 Blood 108 2006 600 608 16569767
- Couture C. Songyang Z. Jascur T. Williams S. Tailor P. Cantley L.C. Mustelin T. Regulation of the Lck SH2 domain by tyrosine phosphorylation J. Biol. Chem. 271 1996 24880 24884 8798764
- Cox J. Mann M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification Nat. Biotechnol. 26 2008 1367 1372 19029910
- Cunnick J.M. Meng S. Ren Y. Desponts C. Wang H.-G. Djeu J.Y. Wu J. Regulation of the mitogen-activated protein kinase signaling pathway by SHP2 J. Biol. Chem. 277 2002 9498 9504 11779868
- de Aós I. Metzger M.H. Exley M. Dahl C.E. Misra S. Zheng D. Varticovski L. Terhorst C. Sancho J. Tyrosine phosphorylation of the CD3-epsilon subunit of the T cell antigen receptor mediates enhanced association with phosphatidylinositol 3-kinase in Jurkat T cells J. Biol. Chem. 272 1997 25310 25318 9312149
- Evdokimova V. Ruzanov P. Anglesio M.S. Sorokin A.V. Ovchinnikov L.P. Buckley J. Triche T.J. Sonenberg N. Sorensen P.H.B. Akt-mediated YB-1 phosphorylation activates translation of silent mRNA species Mol. Cell. Biol. 26 2006 277 292 16354698 PMC1317623
- Giansanti P. Aye T.T. van den Toorn H. Peng M. van Breukelen B. Heck A.J.R. An augmented multiple-protease-based human phosphopeptide atlas Cell Rep. 11 2015 1834 1843 26074081
- Hatakeyama M. Kono T. Kobayashi N. Kawahara A. Levin S.D. Perlmutter R.M. Taniguchi T. Interaction of the IL-2 receptor with the src-family kinase p56lck: identification of novel intermolecular association Science 252 1991 1523 1528 2047859
- Hedrick S.M. Hess Michelini R. Doedens A.L. Goldrath A.W. Stone E.L. FOXO transcription factors throughout T cell biology Nat. Rev. Immunol. 12 2012 649 661 22918467 PMC3875397
- Horak I.D. Gress R.E. Lucas P.J. Horak E.M. Waldmann T.A. Bolen J.B. T-lymphocyte interleukin 2-dependent tyrosine protein kinase signal transduction involves the activation of p56lck Proc. Natl. Acad. Sci. USA 88 1991 1996 2000 2000405 PMC51153
- John S. Robbins C.M. Leonard W.J. An IL-2 response element in the human IL-2 receptor alpha chain promoter is a composite element that binds Stat5, Elf-1, HMG-I(Y) and a GATA family protein EMBO J. 15 1996 5627 5635 8896456 PMC452307
- Kim H.P. Kelly J. Leonard W.J. The basis for IL-2-induced IL-2 receptor alpha chain gene regulation: importance of two widely separated IL-2 response elements Immunity 15 2001 159 172 11485747
- Kobayashi N. Kono T. Hatakeyama M. Minami Y. Miyazaki T. Perlmutter R.M. Taniguchi T. Functional coupling of the src-family protein tyrosine kinases p59fyn and p53/56lyn with the interleukin 2 receptor: implications for redundancy and pleiotropism in cytokine signal transduction Proc. Natl. Acad. Sci. USA 90 1993 4201 4205 8483935 PMC46474
- Lee S. Truesdell S.S. Bukhari S.I.A. Lee J.H. LeTonqueze O. Vasudevan S. Upregulation of eIF5B controls cell-cycle arrest and specific developmental stages Proc. Natl. Acad. Sci. USA 111 2014 E4315 E4322 25261552 PMC4205643
- Liao W. Lin J.-X. Leonard W.J. Interleukin-2 at the crossroads of effector responses, tolerance, and immunotherapy Immunity 38 2013 13 25 23352221 PMC3610532
- Lin J.-X. Li P. Liu D. Jin H.-T. He J. Ata Ur Rasheed M. Rochman Y. Wang L. Cui K. Liu C. Critical Role of STAT5 transcription factor tetramerization for cytokine responses and normal immune function Immunity 36 2012 586 599 22520852 PMC3551341
- Lucas C.L. Kuehn H.S. Zhao F. Niemela J.E. Deenick E.K. Palendira U. Avery D.T. Moens L. Cannons J.L. Biancalana M. Dominant-activating germline mutations in the gene encoding the PI(3)K catalytic subunit p110δ result in T cell senescence and human immunodeficiency Nat. Immunol. 15 2014 88 97 24165795 PMC4209962
- Luo W. Slebos R.J. Hill S. Li M. Brábek J. Amanchy R. Chaerkady R. Pandey A. Ham A.-J.L. Hanks S.K. Global impact of oncogenic Src on a phosphotyrosine proteome J. Proteome Res. 7 2008 3447 3460 18563927 PMC2579752
- Macintyre A.N. Finlay D. Preston G. Sinclair L.V. Waugh C.M. Tamas P. Feijoo C. Okkenhaug K. Cantrell D.A. Protein kinase B controls transcriptional programs that direct cytotoxic T cell fate but is dispensable for T cell metabolism Immunity 34 2011 224 236 21295499 PMC3052433
- Maksumova L. Le H.T. Muratkhodjaev F. Davidson D. Veillette A. Pallen C.J. Protein tyrosine phosphatase alpha regulates Fyn activity and Cbp/PAG phosphorylation in thymocyte lipid rafts J. Immunol. 175 2005 7947 7956 16339530
- Marth J.D. Cooper J.A. King C.S. Ziegler S.F. Tinker D.A. Overell R.W. Krebs E.G. Perlmutter R.M. Neoplastic transformation induced by an activated lymphocyte-specific protein tyrosine kinase (pp56lck) Mol. Cell. Biol. 8 1988 540 550 3352600 PMC363178
- Mitra S. Ring A.M. Amarnath S. Spangler J.B. Li P. Ju W. Fischer S. Oh J. Spolski R. Weiskopf K. Interleukin-2 activity can be fine tuned with engineered receptor signaling clamps Immunity 42 2015 826 838 25992859 PMC4560365
- Mizuno K. Signaling mechanisms and functional roles of cofilin phosphorylation and dephosphorylation Cell. Signal. 25 2013 457 469 23153585
- Najafov A. Shpiro N. Alessi D.R. Akt is efficiently activated by PIF-pocket- and PtdIns(3,4,5)P3-dependent mechanisms leading to resistance to PDK1 inhibitors Biochem. J. 448 2012 285 295 23030823
- Navarro M.N. Goebel J. Feijoo-Carnero C. Morrice N. Cantrell D.A. Phosphoproteomic analysis reveals an intrinsic pathway for the regulation of histone deacetylase 7 that controls the function of cytotoxic T lymphocytes Nat. Immunol. 12 2011 352 361 21399638 PMC3110993
- Navarro M.N. Feijoo-Carnero C. Arandilla A.G. Trost M. Cantrell D.A. Protein kinase D2 is a digital amplifier of T cell receptor-stimulated diacylglycerol signaling in naïve CD8+ T cells Sci. Signal. 7 2014 ra99 25336615 PMC4768351
- Nika K. Soldani C. Salek M. Paster W. Gray A. Etzensperger R. Fugger L. Polzella P. Cerundolo V. Dushek O. Constitutively active Lck kinase in T cells drives antigen receptor signal transduction Immunity 32 2010 766 777 20541955 PMC2996607
- O’Shea J.J. Schwartz D.M. Villarino A.V. Gadina M. McInnes I.B. Laurence A. The JAK-STAT pathway: impact on human disease and therapeutic intervention Annu. Rev. Med. 66 2015 311 328 25587654 PMC5634336
- Okkenhaug K. Ali K. Vanhaesebroeck B. Antigen receptor signalling: a distinctive role for the p110delta isoform of PI3K Trends Immunol. 28 2007 80 87 17208518 PMC2358943
- Osinalde N. Moss H. Arrizabalaga O. Omaetxebarria M.J. Blagoev B. Zubiaga A.M. Fullaondo A. Arizmendi J.M. Kratchmarova I. Interleukin-2 signaling pathway analysis by quantitative phosphoproteomics J. Proteomics 75 2011 177 191 21722762
- Osinalde N. Sanchez-Quiles V. Akimov V. Guerra B. Blagoev B. Kratchmarova I. Simultaneous dissection and comparison of IL-2 and IL-15 signaling pathways by global quantitative phosphoproteomics Proteomics 15 2015 520 531 25142963
- Pipkin M.E. Sacks J.A. Cruz-Guilloty F. Lichtenheld M.G. Bevan M.J. Rao A. Interleukin-2 and inflammation induce distinct transcriptional programs that promote the differentiation of effector cytolytic T cells Immunity 32 2010 79 90 20096607 PMC2906224
- Pircher H. Bürki K. Lang R. Hengartner H. Zinkernagel R.M. Tolerance induction in double specific T-cell receptor transgenic mice varies with antigen Nature 342 1989 559 561 2573841
- Preston G.C. Sinclair L.V. Kaskar A. Hukelmann J.L. Navarro M.N. Ferrero I. MacDonald H.R. Cowling V.H. Cantrell D.A. Single cell tuning of Myc expression by antigen receptor signal strength and interleukin-2 in T lymphocytes EMBO J. 34 2015 2008 2024 26136212 PMC4551349
- Ray J.P. Staron M.M. Shyer J.A. Ho P.-C. Marshall H.D. Gray S.M. Laidlaw B.J. Araki K. Ahmed R. Kaech S.M. Craft J. The interleukin-2-mTORc1 kinase axis defines the signaling, differentiation, and metabolism of T helper 1 and follicular B helper T cells Immunity 43 2015 690 702 26410627 PMC4618086
- Safari F. Murata-Kamiya N. Saito Y. Hatakeyama M. Mammalian Pragmin regulates Src family kinases via the Glu-Pro-Ile-Tyr-Ala (EPIYA) motif that is exploited by bacterial effectors Proc. Natl. Acad. Sci. USA 108 2011 14938 14943 21873224 PMC3169139
- Smith G.A. Uchida K. Weiss A. Taunton J. Essential biphasic role for JAK3 catalytic activity in IL-2 receptor signaling Nat. Chem. Biol. 12 2016 373 379 27018889 PMC4837022
- Spangler J.B. Tomala J. Luca V.C. Jude K.M. Dong S. Ring A.M. Votavova P. Pepper M. Kovar M. Garcia K.C. Antibodies to interleukin-2 elicit selective T cell subset potentiation through distinct conformational mechanisms Immunity 42 2015 815 825 25992858 PMC4439582
- Stone E.L. Pepper M. Katayama C.D. Kerdiles Y.M. Lai C.-Y. Emslie E. Lin Y.C. Yang E. Goldrath A.W. Li M.O. ICOS coreceptor signaling inactivates the transcription factor FOXO1 to promote Tfh cell differentiation Immunity 42 2015 239 251 25692700 PMC4334393
- Tanaka M. Maeda K. Nakashima K. Chicken alpha-enolase but not beta-enolase has a Src-dependent tyrosine-phosphorylation site: cDNA cloning and nucleotide sequence analysis J. Biochem. 117 1995 554 559 7629021
- Titz B. Low T. Komisopoulou E. Chen S.S. Rubbi L. Graeber T.G. The proximal signaling network of the BCR-ABL1 oncogene shows a modular organization Oncogene 29 2010 5895 5910 20697350 PMC5308218
- Vindis C. Teli T. Cerretti D.P. Turner C.E. Huynh-Do U. EphB1-mediated cell migration requires the phosphorylation of paxillin at Tyr-31/Tyr-118 J. Biol. Chem. 279 2004 27965 27970 15107421
- Zhang Z. Shen K. Lu W. Cole P.A. The role of C-terminal tyrosine phosphorylation in the regulation of SHP-1 explored via expressed protein ligation J. Biol. Chem. 278 2003 4668 4674 12468540
- Zhou Y.J. Magnuson K.S. Cheng T.P. Gadina M. Frucht D.M. Galon J. Candotti F. Geahlen R.L. Changelian P.S. O’Shea J.J. Hierarchy of protein tyrosine kinases in interleukin-2 (IL-2) signaling: activation of syk depends on Jak3; however, neither Syk nor Lck is required for IL-2-mediated STAT activation Mol. Cell. Biol. 20 2000 4371 4380 10825200 PMC85804
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- Authors
- Ross, S. H., Rollings, C., Anderson, K. E., Hawkins, P. T., Stephens, L. R., & Cantrell, D. A.
- Affiliations
- University of Dundee (GB)
- Babraham Institute (GB)
- Corresponding author
- Cantrell, Doreen A.
- ORCID
- Venue
- Immunity
- Publisher
- Elsevier BV
- Published
- 2016-09
- Volume
- 45
- Issue
- 3
- Pages
- 685-700
- ISSN
- 1074-7613 (ISSN-L)
- PMC
- PMC5040828
- OpenAlex
- W2512028013
- Cited by
- 89
- References
- 3
- Citations by year
- 2026: 6 · 2025: 7 · 2024: 13 · 2023: 9 · 2022: 9 · 2021: 9 · 2020: 8 · 2019: 8 · 2018: 6 · 2017: 12 · 2016: 2
- FWCI
- 5.03
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Immunity (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Discovery Research Portal (University of Dundee) (submittedVersion, cc-by)
- Funders
- GlaxoSmithKline
- University of Dundee
- Wellcome Trust (097418/Z/11/Z, /Z/11/Z, 097418)
- Tenovus
- Directorate for Biological Sciences
- Biotechnology and Biological Sciences Research Council (BBS/E/B/000C0415, 1252752)
- Topics
- Health Sciences › Medicine › Oncology › Cytokine Signaling Pathways and Interactions
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
- Keywords
- interleukin-2 signaling, CD8+ T cells, cytotoxic T lymphocytes, JAK kinases, protein phosphorylation, phosphoproteomics, Src family kinases, phosphatidylinositol 3,4,5-trisphosphate, Akt signaling, transcription factor phosphorylation, chromatin regulators, vesicle trafficking, cytoskeleton, T cell fate
- MeSH terms
- Actins, Animals, Cell Differentiation, Interleukin-2, Phosphorylation, RNA, Messenger, Signal Transduction, Trans-Activators, Protein Serine-Threonine Kinases, CD8-Positive T-Lymphocytes, Proteome, GTP Phosphohydrolases, Cell Proliferation, Mice, Janus Kinases
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