title: UBP43 (USP18) specifically removes ISG15 from conjugated proteins.
abstract: UBP43 shows significant homology to well characterized ubiquitin-specific proteases and previously was shown to hydrolyze ubiquitin-beta-galactosidase fusions in Escherichia coli. In our assays, the activity of UBP43 toward Ub fusions was undetectable in vitro directing us to investigate the possibility of Ub-like proteins such as SUMO, Nedd8, and ISG15 as probable substrates. We consequently demonstrate that UBP43 can efficiently cleave only ISG15 fusions including native ISG15 conjugates linked via isopeptide bonds. In addition to commonly used methods we introduce a new experimental design featuring ISG15-UBP43 fusion self-processing. Deletion of the UBP43 gene in mouse leads to a massive increase of ISG15 conjugates in tissues indicating that UBP43 is a major ISG15-specific protease. UBP43 is the first bona fide ISG15-specific protease reported. Both ISG15 and UBP43 genes are known to be strongly induced by interferon, genotoxic stress, and viral infection. We postulate that UBP43 is necessary to maintain a critical cellular balance of ISG15-conjugated proteins in both healthy and stressed organisms.
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Malakhov
authors.forename: Oxana A
authors.initials: OA
authors.lastname: Malakhova
authors.forename: Keun Il
authors.initials: KI
authors.lastname: Kim
authors.forename: Kenneth J
authors.initials: KJ
authors.lastname: Ritchie
authors.forename: Dong-Er
authors.initials: DE
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D008168:Lung / Q000378:metabolism; D051379:Mice; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D013379:Substrate Specificity; D043222:Ubiquitin Thiolesterase; D014452:Ubiquitins / Q000031:analogs & derivatives*
chemical_list: D016207:Cytokines; D018076:DNA, Complementary; C489550:G1p2 protein, mouse; D014452:Ubiquitins; C074034:ISG15 protein, human; D010450:Endopeptidases; C118306:Usp18 protein, mouse; C485560:USP18 protein, human; D043222:Ubiquitin Thiolesterase