The proteomic analysis of endogenous FAT10 substrates identifies p62/SQSTM1 as a substrate of FAT10ylation

Annette Aichem, Birte Kalveram, Valentina Spinnenhirn, Kathrin Kluge, Nicola Catone, Terje Johansen, Marcus Groettrup

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

FAT10 is a ubiquitin-like modifier proposed to function in apoptosis induction, cell cycle control and NF-κB activation. Upon induction by pro-inflammatory cytokines, hundreds of endogenous substrates become covalently conjugated to FAT10 leading to their proteasomal degradation. Nevertheless, only three substrates have been identified so far to which FAT10 becomes covalently attached through a non-reducible isopeptide bond, and these are the FAT10-conjugating enzyme USE1 which auto-FAT10ylates itself in cis, the tumor suppressor p53 and the ubiquitin-activating enzyme UBE1 (UBA1). To identify additional FAT10 substrates and interaction partners, we used a new monoclonal FAT10-specific antibody to immunopurify endogenous FAT10 conjugates from interferon (IFN)γ- and tumor necrosis factor (TNF)α-stimulated cells for identification by mass spectrometry. In addition to two already known FAT10-interacting proteins, histone deacetylase 6 and UBA6, we identified 569 novel FAT10-interacting proteins involved in different functional pathways such as autophagy, cell cycle regulation, apoptosis and cancer. Thirty-one percent of all identified proteins were categorized as putative covalently linked substrates. One of the identified proteins, the autophagosomal receptor p62/SQSTM1, was further investigated. p62 becomes covalently mono-FAT10ylated at several lysines, and FAT10 colocalizes with p62 in p62 bodies. Strikingly, FAT10ylation of p62 leads to its proteasomal degradation, and prolonged induction of endogenous FAT10 expression by proinflammatory cytokines leads to a decrease of endogenous p62. The elucidation of the FAT10 degradome should enable a better understanding of why FAT10 has evolved as an additional transferable tag for proteasomal degradation.

Original languageEnglish (US)
Pages (from-to)4576-4585
Number of pages10
JournalJournal of Cell Science
Volume125
Issue number19
DOIs
StatePublished - 2012
Externally publishedYes

Fingerprint

Proteomics
Proteins
Ubiquitin-Activating Enzymes
Apoptosis
Cytokines
Histone Deacetylases
Autophagy
Ubiquitin
Cell Cycle Checkpoints
Interferons
Lysine
Mass Spectrometry
Neoplasms
Cell Cycle
Tumor Necrosis Factor-alpha
Antibodies
Enzymes

Keywords

  • FAT10
  • p62
  • Proteasome
  • Proteomics
  • Ubiquitin

ASJC Scopus subject areas

  • Cell Biology

Cite this

Aichem, A., Kalveram, B., Spinnenhirn, V., Kluge, K., Catone, N., Johansen, T., & Groettrup, M. (2012). The proteomic analysis of endogenous FAT10 substrates identifies p62/SQSTM1 as a substrate of FAT10ylation. Journal of Cell Science, 125(19), 4576-4585. https://doi.org/10.1242/jcs.107789

The proteomic analysis of endogenous FAT10 substrates identifies p62/SQSTM1 as a substrate of FAT10ylation. / Aichem, Annette; Kalveram, Birte; Spinnenhirn, Valentina; Kluge, Kathrin; Catone, Nicola; Johansen, Terje; Groettrup, Marcus.

In: Journal of Cell Science, Vol. 125, No. 19, 2012, p. 4576-4585.

Research output: Contribution to journalArticle

Aichem, A, Kalveram, B, Spinnenhirn, V, Kluge, K, Catone, N, Johansen, T & Groettrup, M 2012, 'The proteomic analysis of endogenous FAT10 substrates identifies p62/SQSTM1 as a substrate of FAT10ylation', Journal of Cell Science, vol. 125, no. 19, pp. 4576-4585. https://doi.org/10.1242/jcs.107789
Aichem, Annette ; Kalveram, Birte ; Spinnenhirn, Valentina ; Kluge, Kathrin ; Catone, Nicola ; Johansen, Terje ; Groettrup, Marcus. / The proteomic analysis of endogenous FAT10 substrates identifies p62/SQSTM1 as a substrate of FAT10ylation. In: Journal of Cell Science. 2012 ; Vol. 125, No. 19. pp. 4576-4585.
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