title: Negative regulation of ISG15 E3 ligase EFP through its autoISGylation.
abstract: The function of ubiquitin-like protein ISG15 and protein modification by ISG15 (ISGylation) has been an enigma for many years. Recently, the research of ISGylation has been accelerated by the identification of the enzymes involved in the ISG15 conjugation process. Our previous study identified the interferon inducible protein EFP as an ISG15 isopeptide ligase (E3) for 14-3-3sigma. In this study, we show that ISG15 E3 ligase EFP can be modified by ISG15. Two ubiquitin E2 conjugating enzymes, UbcH6 and UbcH8, can support ISGylation of EFP. The Ring-finger domain of EFP is important for its ISGylation. Full-length EFP can enhance the ISGylation of Ring domain deleted EFP, indicating EFP can function as an ISG15 E3 ligase for itself. We also determined the ISGylation site of EFP and created its ISGylation resistant mutant EFP-K117R. Compared to the wild-type EFP, this mutant further increases the ISGylation of 14-3-3sigma. Thus we propose that autoISGylation of EFP negatively regulates its ISG15 E3 ligase activity for 14-3-3sigma.
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Weiguo
authors.initials: W
authors.lastname: Zou
authors.forename: Ji
authors.initials: J
authors.lastname: Wang
authors.forename: Dong-Er
authors.initials: DE
authors.lastname: Zhang
mesh_terms: D001665:Binding Sites; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism*; D004789:Enzyme Activation; D005246:Feedback / Q000502:physiology; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014157:Transcription Factors / Q000378:metabolism*; D000070584:Tripartite Motif Proteins; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D014452:Ubiquitins / Q000378:metabolism*
chemical_list: D016207:Cytokines; D014157:Transcription Factors; D000070584:Tripartite Motif Proteins; D014452:Ubiquitins; C074034:ISG15 protein, human; C084523:TRIM25 protein, human; D044767:Ubiquitin-Protein Ligases