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Phosphoproteomic analysis reveals an intrinsic pathway for the regulation of histone deacetylase 7 that controls the function of cytotoxic T lymphocytes

Nature Immunology volume 12, pages 352–361 (2011)Cite this article

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Abstract

Here we report an unbiased analysis of the cytotoxic T lymphocyte (CTL) serine-threonine phosphoproteome by high-resolution mass spectrometry. We identified approximately 2,000 phosphorylations in CTLs, of which approximately 450 were controlled by T cell antigen receptor (TCR) signaling. A significantly overrepresented group of molecules identified included transcription activators, corepressors and chromatin regulators. A focus on chromatin regulators showed that CTLs had high expression of the histone deacetylase HDAC7 but continually phosphorylated and exported this transcriptional repressor from the nucleus. Dephosphorylation of HDAC7 resulted in its accumulation in the nucleus and suppressed expression of genes encoding key cytokines, cytokine receptors and adhesion molecules that determine CTL function. Screening of the CTL phosphoproteome has thus identified intrinsic pathways of serine-threonine phosphorylation that target chromatin regulators and determine the CTL functional program.

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Figure 1: Analysis of the basal and TCR-regulated phosphoproteome in CTLs.
Figure 2: Identification of consistent phosphorylations in CTLs by Ingenuity Pathway Analysis software.
Figure 3: Phosphorylated chromatin regulators in CTLs.
Figure 4: Subcellular distribution of HDAC7 in CTLs.
Figure 5: Exclusion of HDAC7 from the nucleus is required for normal CTL function.
Figure 6: Exclusion of HDAC7 from the nucleus is required for expression of the high-affinity IL-2 receptor.

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Acknowledgements

We thank members of the Biological Services Unit, R. Clarke of the Flow Cytometry Facility and members of the Cantrell laboratory for critical reading of the manuscript; and the Finnish DNA Microarray Centre at the Centre for Biotechnology (Turku, Finland) and N. Schurch for microarray analysis. Supported by the Wellcome Trust (065975/Z/01/A) and the Medical Research Council (J.G.).

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  1. Division of Immunology and Cell Biology, The College of Life Sciences, The University of Dundee, Dundee, Scotland, UK

    Maria N Navarro, Jurgen Goebel, Carmen Feijoo-Carnero & Doreen A Cantrell

  2. The Beatson Institute for Cancer Research, Glasgow, Scotland, UK

    Nick Morrice

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Contributions

M.N.N., J.G. and C.F.-C. did the experiments and analyzed the results; N.M. supervised SILAC methodology and bioinformatic analysis; and M.N.N. and D.A.C. designed the experiments, analyzed the results and wrote the paper.

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Correspondence to Doreen A Cantrell.

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Navarro, M., Goebel, J., Feijoo-Carnero, C. et al. Phosphoproteomic analysis reveals an intrinsic pathway for the regulation of histone deacetylase 7 that controls the function of cytotoxic T lymphocytes. Nat Immunol 12, 352–361 (2011). https://doi.org/10.1038/ni.2008

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