title: Conjugation of the ubiquitin activating enzyme UBE1 with the ubiquitin-like modifier FAT10 targets it for proteasomal degradation.
abstract: The ubiquitin-like modifier HLA-F adjacent transcript 10 (FAT10) directly targets its substrates for proteasomal degradation by becoming covalently attached via its C-terminal diglycine motif to internal lysine residues of its substrate proteins. The conjugation machinery consists of the bispecific E1 activating enzyme Ubiquitin-like modifier activating enzyme 6 (UBA6), the likewise bispecific E2 conjugating enzyme UBA6-specific E2 enzyme 1 (USE1), and possibly E3 ligases. By mass spectrometry analysis the ubiquitin E1 activating enzyme ubiquitin-activating enzyme 1 (UBE1) was identified as putative substrate of FAT10. Here, we confirm that UBE1 and FAT10 form a stable non-reducible conjugate under overexpression as well as under endogenous conditions after induction of endogenous FAT10 expression with proinflammatory cytokines. FAT10ylation of UBE1 depends on the diglycine motif of FAT10. By specifically downregulating FAT10, UBA6 or USE1 with siRNAs, we show that UBE1 modification depends on the FAT10 conjugation pathway. Furthermore, we confirm that UBE1 does not act as a second E1 activating enzyme for FAT10 but that FAT10ylation of UBE1 leads to its proteasomal degradation, implying a putative regulatory role of FAT10 in the ubiquitin conjugation pathway.
authors.affiliation: Biotechnology Institute Thurgau at the University of Konstanz, Unterseestrasse 47, CH-8280, Kreuzlingen, Switzerland; Division of Immunology, Department of Biology, University of Konstanz, D-78457, Konstanz, Germany.
authors.forename: Johanna
authors.initials: J
authors.lastname: Bialas
authors.affiliation: Biotechnology Institute Thurgau at the University of Konstanz, Unterseestrasse 47, CH-8280, Kreuzlingen, Switzerland; Division of Immunology, Department of Biology, University of Konstanz, D-78457, Konstanz, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Groettrup
authors.affiliation: Biotechnology Institute Thurgau at the University of Konstanz, Unterseestrasse 47, CH-8280, Kreuzlingen, Switzerland.
authors.forename: Annette
authors.initials: A
authors.lastname: Aichem
mesh_terms: D006801:Humans; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011485:Protein Binding; D059748:Proteolysis; D044764:Ubiquitin-Activating Enzymes / Q000737:chemistry / Q000378:metabolism*; D054875:Ubiquitination; D014452:Ubiquitins / Q000737:chemistry / Q000378:metabolism*
chemical_list: C471538:UBA1 protein, human; C109184:UBD protein, human; D014452:Ubiquitins; D046988:Proteasome Endopeptidase Complex; D044764:Ubiquitin-Activating Enzymes