Structural basis and specificity of human otubain 1-mediated deubiquitination

Mariola J. Edelmann, Alexander Iphöfer, Masato Akutsu, Mikael Altun, Katalin di Gleria, Holger B. Kramer, Edda Fiebiger, Sirano Dhe-Paganon, Benedikt M. Kessler

Research output: Contribution to journalArticlepeer-review

125 Citations (Scopus)

Abstract

OTUB (otubain) 1 is a human deubiquitinating enzyme that is implicated in mediating lymphocyte antigen responsiveness, but whose molecular function is generally not well defined. A structural analysis of OTUB1 shows differences in accessibility to the active site and in surface properties of the substrate-binding regions when compared with its close homologue, OTUB2, suggesting variations in regulatory mechanisms and substrate specificity. Biochemical analysis reveals that OTUB1 has a preference for cleaving Lys48-linked polyubiquitin chains over Lys63-linked polyubiquitin chains, and it is capable of cleaving NEDD8 (neural-precursor-cell-expressed developmentally down-regulated 8), but not SUMO (small ubiquitin-related modifier) 1/2/3 and ISG15 (interferon-stimulated gene 15) conjugates. A functional comparison of OTUB1 and OTUB2 indicated a differential reactivity towards ubiquitin-based active-site probes carrying a vinyl methyl ester, a 2-chloroethyl or a 2-bromoethyl group at the C-terminus. Mutational analysis suggested that a narrow P1′ site, as observed in OTUB1, correlates with its ability to preferentially cleave Lys48-linked ubiquitin chains. Analysis of cellular interaction partners of OTUB1 by co-immunoprecipitation and MS/MS (tandem mass spectrometry) experiments demonstrated that FUS [fusion involved in t(12;6) in malignant liposarcoma; also known as TLS (translocation in liposarcoma) or CHOP (CCAAT/enhancer-binding protein homologous protein)] and RACK1 [receptor for activated kinase 1; also known as GNB2L1 (guanine-nucleotide-binding protein β polypeptide 2-like 1)] are part of OTUB1-containing complexes, pointing towards a molecular function of this deubiquitinating enzyme in RNA processing and cell adhesion/morphology.

Original languageEnglish
Pages (from-to)379-390
Number of pages12
JournalBiochemical Journal
Volume418
Issue number2
DOIs
Publication statusPublished - 2009 Mar 1
Externally publishedYes

Keywords

  • Deubiquitinating enzyme (DUB)
  • Interferon-stimulated gene 15 (ISG15)
  • Neural-precursor-cell-expressed developmentally down-regulated 8 (NEDD8)
  • Otubain 1 (OTUB1)
  • Small ubiquitin-related modifier (SUMO)
  • Ubiquitin

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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