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Uncovering the SUMOylation and ubiquitylation crosstalk in human cells using sequential peptide immunopurification

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  • Frédéric Lamoliatte

    (Institute for Research in Immunology and Cancer, Université de Montréal
    Université de Montréal)

  • Francis P. McManus

    (Institute for Research in Immunology and Cancer, Université de Montréal)

  • Ghizlane Maarifi

    (INSERM UMR-S1124, Université Paris Descartes)

  • Mounira K. Chelbi-Alix

    (INSERM UMR-S1124, Université Paris Descartes)

  • Pierre Thibault

    (Institute for Research in Immunology and Cancer, Université de Montréal
    Université de Montréal
    Université de Montréal)

Abstract

Crosstalk between the SUMO and ubiquitin pathways has recently been reported. However, no approach currently exists to determine the interrelationship between these modifications. Here, we report an optimized immunoaffinity method that permits the study of both protein ubiquitylation and SUMOylation from a single sample. This method enables the unprecedented identification of 10,388 SUMO sites in HEK293 cells. The sequential use of SUMO and ubiquitin remnant immunoaffinity purification facilitates the dynamic profiling of SUMOylated and ubiquitylated proteins in HEK293 cells treated with the proteasome inhibitor MG132. Quantitative proteomic analyses reveals crosstalk between substrates that control protein degradation, and highlights co-regulation of SUMOylation and ubiquitylation levels on deubiquitinase enzymes and the SUMOylation of proteasome subunits. The SUMOylation of the proteasome affects its recruitment to promyelocytic leukemia protein (PML) nuclear bodies, and PML lacking the SUMO interacting motif fails to colocalize with SUMOylated proteasome further demonstrating that this motif is required for PML catabolism.

Suggested Citation

  • Frédéric Lamoliatte & Francis P. McManus & Ghizlane Maarifi & Mounira K. Chelbi-Alix & Pierre Thibault, 2017. "Uncovering the SUMOylation and ubiquitylation crosstalk in human cells using sequential peptide immunopurification," Nature Communications, Nature, vol. 8(1), pages 1-11, April.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms14109
    DOI: 10.1038/ncomms14109
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