{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,3]],"date-time":"2026-11-03T19:11:16Z","timestamp":1793733076706,"version":"4.6.1"},"reference-count":28,"publisher":"National Academy of Sciences","issue":"6","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2011,2,8]]},"abstract":"<jats:p>Crimean Congo hemorrhagic fever virus (CCHFV) is a deadly human pathogen that evades innate immune responses by efficiently interfering with antiviral signaling pathways mediated by NF-\u03baB, IRF3, and IFN\u03b1\/\u03b2. These pathways rely on protein ubiquitination for their activation, and one outcome is the modification of proteins with the ubiquitin (Ub)-like modifier interferon-stimulated gene (ISG)15. CCHFV and related viruses encode a deubiquitinase (DUB) of the ovarian tumor (OTU) family, which unlike eukaryotic OTU DUBs also targets ISG15 modifications. Here we characterized the viral OTU domain of CCHFV (vOTU) biochemically and structurally, revealing that it hydrolyzes four out of six tested Ub linkages, but lacks activity against linear and K29-linked Ub chains. vOTU cleaved Ub and ISG15 with similar kinetics, and we were able to understand vOTU cross-reactivity at the molecular level from crystal structures of vOTU in complex with Ub and ISG15. An N-terminal extension in vOTU not present in eukaryotic OTU binds to the hydrophobic Ile44 patch of Ub, which results in a dramatically different Ub orientation compared to a eukaryotic OTU\u2013Ub complex. The C-terminal Ub-like fold of ISG15 (ISG15-C) adopts an equivalent binding orientation. Interestingly, ISG15-C contains an additional second hydrophobic surface that is specifically contacted by vOTU. These subtle differences in Ub\/ISG15 binding allowed the design of vOTU variants specific for either Ub or ISG15, which will be useful tools to understand the relative contribution of ubiquitination vs. ISGylation in viral infection. Furthermore, the crystal structures will allow structure-based design of antiviral agents targeting this enzyme.<\/jats:p>","DOI":"10.1073\/pnas.1015287108","type":"journal-article","created":{"date-parts":[[2011,1,25]],"date-time":"2011-01-25T22:47:34Z","timestamp":1295995654000},"page":"2228-2233","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":156,"title":["Molecular basis for ubiquitin and ISG15 cross-reactivity in viral ovarian tumor domains"],"prefix":"10.1073","volume":"108","author":[{"given":"Masato","family":"Akutsu","sequence":"first","affiliation":[{"name":"Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Ye","sequence":"additional","affiliation":[{"name":"Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satpal","family":"Virdee","sequence":"additional","affiliation":[{"name":"Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason W.","family":"Chin","sequence":"additional","affiliation":[{"name":"Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Komander","sequence":"additional","affiliation":[{"name":"Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2011,1,25]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1016\/j.cell.2006.02.015","article-title":"Pathogen recognition and innate immunity","volume":"124","author":"Akira S","year":"2006","unstructured":"S Akira, S Uematsu, O Takeuchi, Pathogen recognition and innate immunity. Cell 124, 783\u2013801 (2006).","journal-title":"Cell"},{"key":"e_1_3_4_2_2","doi-asserted-by":"crossref","first-page":"R328","DOI":"10.1016\/j.cub.2010.01.044","article-title":"Sensing and signaling in antiviral innate immunity","volume":"20","author":"O\u2019Neill LA","year":"2010","unstructured":"LA O\u2019Neill, AG Bowie, Sensing and signaling in antiviral innate immunity. Curr Biol 20, R328\u2013333 (2010).","journal-title":"Curr Biol"},{"key":"e_1_3_4_3_2","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1038\/nri2314","article-title":"Interferon-inducible antiviral effectors","volume":"8","author":"Sadler AJ","year":"2008","unstructured":"AJ Sadler, BRG Williams, Interferon-inducible antiviral effectors. Nat Rev Immunol 8, 559\u2013568 (2008).","journal-title":"Nat Rev Immunol"},{"key":"e_1_3_4_4_2","doi-asserted-by":"crossref","first-page":"937","DOI":"10.1042\/BST0370937","article-title":"The emerging complexity of protein ubiquitination","volume":"37","author":"Komander D","year":"2009","unstructured":"D Komander, The emerging complexity of protein ubiquitination. Biochem Soc Trans 37, 937\u2013953 (2009).","journal-title":"Biochem Soc Trans"},{"key":"e_1_3_4_5_2","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/j.cell.2010.03.029","article-title":"Reconstitution of the RIG-I pathway reveals a signaling role of unanchored polyubiquitin chains in innate immunity","volume":"141","author":"Zeng W","year":"2010","unstructured":"W Zeng, et al., Reconstitution of the RIG-I pathway reveals a signaling role of unanchored polyubiquitin chains in innate immunity. Cell 141, 315\u2013330 (2010).","journal-title":"Cell"},{"key":"e_1_3_4_6_2","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1146\/annurev.biochem.78.070907.102750","article-title":"The role of ubiquitin in NF-kappaB regulatory pathways","volume":"78","author":"Skaug B","year":"2009","unstructured":"B Skaug, X Jiang, ZJ Chen, The role of ubiquitin in NF-kappaB regulatory pathways. Annu Rev Biochem 78, 769\u2013796 (2009).","journal-title":"Annu Rev Biochem"},{"key":"e_1_3_4_7_2","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1038\/ncb1821","article-title":"Involvement of linear polyubiquitylation of NEMO in NF-kappaB activation","volume":"11","author":"Tokunaga F","year":"2009","unstructured":"F Tokunaga, et al., Involvement of linear polyubiquitylation of NEMO in NF-kappaB activation. Nat Cell Biol 11, 123\u2013132 (2009).","journal-title":"Nat Cell Biol"},{"key":"e_1_3_4_8_2","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1038\/nrm2731","article-title":"Breaking the chains: Structure and function of the deubiquitinases","volume":"10","author":"Komander D","year":"2009","unstructured":"D Komander, MJ Clague, S Urb\u00e9, Breaking the chains: Structure and function of the deubiquitinases. Nat Rev Mol Cell Biol 10, 550\u2013563 (2009).","journal-title":"Nat Rev Mol Cell Biol"},{"key":"e_1_3_4_9_2","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1038\/nature02794","article-title":"De-ubiquitination and ubiquitin ligase domains of A20 downregulate NF-kappaB signalling","volume":"430","author":"Wertz I","year":"2004","unstructured":"I Wertz, et al., De-ubiquitination and ubiquitin ligase domains of A20 downregulate NF-kappaB signalling. Nature 430, 694\u2013699 (2004).","journal-title":"Nature"},{"key":"e_1_3_4_10_2","doi-asserted-by":"crossref","first-page":"1628","DOI":"10.1126\/science.1145918","article-title":"DUBA: A deubiquitinase that regulates type I interferon production","volume":"318","author":"Kayagaki N","year":"2007","unstructured":"N Kayagaki, et al., DUBA: A deubiquitinase that regulates type I interferon production. Science 318, 1628\u20131632 (2007).","journal-title":"Science"},{"key":"e_1_3_4_11_2","doi-asserted-by":"crossref","first-page":"2701","DOI":"10.2741\/1730","article-title":"ISG15: A ubiquitin-like enigma","volume":"10","author":"Dao CT","year":"2005","unstructured":"CT Dao, DE Zhang, ISG15: A ubiquitin-like enigma. Front Biosci 10, 2701\u20132722 (2005).","journal-title":"Front Biosci"},{"key":"e_1_3_4_12_2","doi-asserted-by":"crossref","first-page":"27356","DOI":"10.1074\/jbc.M502814200","article-title":"Crystal structure of the interferon-induced ubiquitin-like protein ISG15","volume":"280","author":"Narasimhan J","year":"2005","unstructured":"J Narasimhan, et al., Crystal structure of the interferon-induced ubiquitin-like protein ISG15. J Biol Chem 280, 27356\u201327365 (2005).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_13_2","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/j.cell.2010.09.033","article-title":"Emerging role of ISG15 in antiviral immunity","volume":"143","author":"Skaug B","year":"2010","unstructured":"B Skaug, ZJ Chen, Emerging role of ISG15 in antiviral immunity. Cell 143, 187\u2013190 (2010).","journal-title":"Cell"},{"key":"e_1_3_4_14_2","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1016\/j.molcel.2010.05.002","article-title":"The ISG15 conjugation system broadly targets newly synthesized proteins: implications for the antiviral function of ISG15","volume":"38","author":"Durfee LA","year":"2010","unstructured":"LA Durfee, N Lyon, K Seo, JM Huibregtse, The ISG15 conjugation system broadly targets newly synthesized proteins: implications for the antiviral function of ISG15. Mol Cell 38, 722\u2013732 (2010).","journal-title":"Mol Cell"},{"key":"e_1_3_4_15_2","doi-asserted-by":"crossref","first-page":"527","DOI":"10.1038\/ncb0509-527","article-title":"Viral avoidance and exploitation of the ubiquitin system","volume":"11","author":"Randow F","year":"2009","unstructured":"F Randow, PJ Lehner, Viral avoidance and exploitation of the ubiquitin system. Nat Cell Biol 11, 527\u2013534 (2009).","journal-title":"Nat Cell Biol"},{"key":"e_1_3_4_16_2","doi-asserted-by":"crossref","first-page":"404","DOI":"10.1016\/j.chom.2007.09.014","article-title":"Ovarian tumor domain-containing viral proteases evade ubiquitin- and ISG15-dependent innate immune responses","volume":"2","author":"Frias-Staheli N","year":"2007","unstructured":"N Frias-Staheli, et al., Ovarian tumor domain-containing viral proteases evade ubiquitin- and ISG15-dependent innate immune responses. Cell Host Microbe 2, 404\u2013416 (2007).","journal-title":"Cell Host Microbe"},{"key":"e_1_3_4_17_2","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1016\/j.cytogfr.2008.11.001","article-title":"Interferon and cytokine responses to Crimean Congo hemorrhagic fever virus; an emerging and neglected viral zonoosis","volume":"19","author":"Weber F","year":"2008","unstructured":"F Weber, A Mirazimi, Interferon and cytokine responses to Crimean Congo hemorrhagic fever virus; an emerging and neglected viral zonoosis. Cytokine Growth F R 19, 395\u2013404 (2008).","journal-title":"Cytokine Growth F R"},{"key":"e_1_3_4_18_2","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1038\/nsmb.1873","article-title":"Lys11-linked ubiquitin chains adopt compact conformations and are preferentially hydrolyzed by the deubiquitinase Cezanne","volume":"17","author":"Bremm A","year":"2010","unstructured":"A Bremm, SM Freund, D Komander, Lys11-linked ubiquitin chains adopt compact conformations and are preferentially hydrolyzed by the deubiquitinase Cezanne. Nat Struct Mol Biol 17, 939\u2013947 (2010).","journal-title":"Nat Struct Mol Biol"},{"key":"e_1_3_4_19_2","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1038\/nchembio.426","article-title":"Engineered diubiquitin synthesis reveals Lys29-isopeptide specificity of an OTU deubiquitinase","volume":"6","author":"Virdee S","year":"2010","unstructured":"S Virdee, Y Ye, DP Nguyen, D Komander, JW Chin, Engineered diubiquitin synthesis reveals Lys29-isopeptide specificity of an OTU deubiquitinase. Nat Chem Biol 6, 750\u2013757 (2010).","journal-title":"Nat Chem Biol"},{"key":"e_1_3_4_20_2","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1038\/embor.2009.55","article-title":"Molecular discrimination of structurally equivalent Lys 63-linked and linear polyubiquitin chains","volume":"10","author":"Komander D","year":"2009","unstructured":"D Komander, et al., Molecular discrimination of structurally equivalent Lys 63-linked and linear polyubiquitin chains. 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Chem Biol 9, 1149\u20131159 (2002).","journal-title":"Chem Biol"},{"key":"e_1_3_4_25_2","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1099\/0022-1317-81-4-853","article-title":"Virus-encoded proteinases and proteolytic processing inthe Nidovirales","volume":"81","author":"Ziebuhr J","year":"2000","unstructured":"J Ziebuhr, EJ Snijder, AE Gorbalenya, Virus-encoded proteinases and proteolytic processing inthe Nidovirales. J Gen Virol 81, 853\u2013879 (2000).","journal-title":"J Gen Virol"},{"key":"e_1_3_4_26_2","doi-asserted-by":"crossref","first-page":"9976","DOI":"10.1074\/jbc.M109078200","article-title":"UBP43 (USP18) specifically removes ISG15 from conjugated proteins","volume":"277","author":"Malakhov MP","year":"2002","unstructured":"MP Malakhov, OA Malakhova, KI Kim, KJ Ritchie, DE Zhang, UBP43 (USP18) specifically removes ISG15 from conjugated proteins. J Biol Chem 277, 9976\u20139981 (2002).","journal-title":"J Biol Chem"},{"key":"e_1_3_4_27_2","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.abb.2007.07.006","article-title":"Selectivity in ISG15 and ubiquitin recognition by the SARS coronavirus papain-like protease","volume":"466","author":"Lindner HA","year":"2007","unstructured":"HA Lindner, et al., Selectivity in ISG15 and ubiquitin recognition by the SARS coronavirus papain-like protease. Arch Biochem Biophys 466, 8\u201314 (2007).","journal-title":"Arch Biochem Biophys"},{"key":"e_1_3_4_28_2","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1128\/JVI.01859-09","article-title":"Crimean-Congo hemorrhagic fever virus-encoded ovarian tumor protease activity is dispensable for virus RNA polymerase function","volume":"84","author":"Bergeron E","year":"2009","unstructured":"E Bergeron, CG Albarino, ML Khristova, ST Nichol, Crimean-Congo hemorrhagic fever virus-encoded ovarian tumor protease activity is dispensable for virus RNA polymerase function. J Virol 84, 216\u2013226 (2009).","journal-title":"J Virol"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.1015287108","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,6,7]],"date-time":"2022-06-07T04:17:02Z","timestamp":1654575422000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.1015287108"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,1,25]]},"references-count":28,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2011,2,8]]}},"alternative-id":["10.1073\/pnas.1015287108"],"URL":"https:\/\/doi.org\/10.1073\/pnas.1015287108","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,1,25]]},"assertion":[{"value":"2011-01-25","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}