title: NEDD8 ultimate buster-1L interacts with the ubiquitin-like protein FAT10 and accelerates its degradation.
abstract: FAT10 is an interferon-gamma-inducible ubiquitin-like protein that consists of two ubiquitin-like domains. FAT10 bears a diglycine motif at its C terminus that can form isopeptide bonds to so far unidentified target proteins. Recently we found that FAT10 and its conjugates are rapidly degraded by the proteasome and that the N-terminal fusion of FAT10 to a long lived protein markedly reduces its half-life. FAT10 may hence direct target proteins to the proteasome for degradation. In this study we report a new interaction partner of FAT10 that may link FAT10 to the proteasome. A yeast two-hybrid screen identified NEDD8 ultimate buster-1L (NUB1L) as a non-covalent binding partner of FAT10, and this interaction was confirmed by coimmunoprecipitation and glutathione S-transferase pull-down experiments. NUB1L is also an interferon-inducible protein that has been reported to interact with the ubiquitin-like protein NEDD8, thus leading to accelerated NEDD8 degradation. Here we show that NUB1L binds to FAT10 much stronger than to NEDD8 and that NEDD8 cannot compete with FAT10 for NUB1L binding. The interaction of FAT10 and NUB1L is specific as green fluorescent fusion proteins containing ubiquitin or SUMO-1 do not bind to NUB1L. The coexpression of NUB1L enhanced the degradation rate of FAT10 8-fold, whereas NEDD8 degradation was only accelerated 2-fold. Because NUB1 was shown to bind to the proteasome subunit RPN10 in vitro and to be contained in 26 S proteasome preparations, it may function as a linker that targets FAT10 for degradation by the proteasome.
authors.affiliation: Department of Biology, Division of Immunology, University of Constance, Universitätsstrasse 10, D-78457 Konstanz, Germany.
authors.forename: Mark Steffen
authors.initials: MS
authors.lastname: Hipp
authors.forename: Shahri
authors.initials: S
authors.lastname: Raasi
authors.forename: Marcus
authors.initials: M
authors.lastname: Groettrup
authors.forename: Gunter
authors.initials: G
authors.lastname: Schmidtke
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D003546:Cysteine Endopeptidases / Q000378:metabolism; D015536:Down-Regulation; D006801:Humans; D051379:Mice; D009097:Multienzyme Complexes / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D011485:Protein Binding; D014157:Transcription Factors / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D014452:Ubiquitins / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D009097:Multienzyme Complexes; C430242:NUB1 protein, human; C486597:Nub1 protein, mouse; D014157:Transcription Factors; D014452:Ubiquitins; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex