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HERC5 and the Isgylation Pathway: Critical Modulators of the Antiviral Immune Response

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HERC5 and the Isgylation Pathway: Critical Modulators of the Antiviral Immune Response viruses Review HERC5 and the ISGylation Pathway: Critical Modulators of the Antiviral Immune Response Nicholas A. Mathieu 1, Ermela Paparisto 2, Stephen D. Barr 2 and Donald E. Spratt 1,* 1 Gustaf H. Carlson School of Chemistry and Biochemistry, Clark University, 950 Main St., Worcester, MA 01610, USA; [email protected] 2 Department of Microbiology and Immunology, Schulich School of Medicine and Dentistry, Western University, 1151 Richmond St., London, ON N6A 5C1, Canada; [email protected] (E.P.); [email protected] (S.D.B.) * Correspondence: [email protected]; Tel.: +1-508-793-7536 Abstract: Mammalian cells have developed an elaborate network of immunoproteins that serve to identify and combat viral pathogens. Interferon-stimulated gene 15 (ISG15) is a 15.2 kDa tandem ubiquitin-like protein (UBL) that is used by specific E1–E2–E3 ubiquitin cascade enzymes to interfere with the activity of viral proteins. Recent biochemical studies have demonstrated how the E3 ligase HECT and RCC1-containing protein 5 (HERC5) regulates ISG15 signaling in response to hepatitis C (HCV), influenza-A (IAV), human immunodeficiency virus (HIV), SARS-CoV-2 and other viral infections. Taken together, the potent antiviral activity displayed by HERC5 and ISG15 make them promising drug targets for the development of novel antiviral therapeutics that can augment the host antiviral response. In this review, we examine the emerging role of ISG15 in antiviral immunity with a particular focus on how HERC5 orchestrates the specific and timely ISGylation of viral proteins in response to infection. Citation: Mathieu, N.A.; Paparisto, Keywords: ISG15; ISGylation; HECT E3 ubiquitin ligase; HERC5; antiviral immune response; inter- E.; Barr, S.D.; Spratt, D.E. HERC5 and feron the ISGylation Pathway: Critical Modulators of the Antiviral Immune Response. Viruses 2021, 13, 1102. https://doi.org/10.3390/v13061102 1. Introduction Viral replication is a highly coordinated process that relies heavily on the host cellular Academic Editor: Ricardo Rajsbaum machinery and can disrupt critical cellular activities leading to disease and/or death [1–4]. Viruses exert immense evolutionary pressure on their hosts, driving the development of Received: 1 May 2021 complex cellular immune responses that work to identify, combat and eliminate infectious Accepted: 4 June 2021 pathogens. For example, various mammalian tissues contain immune cells that use pattern Published: 9 June 2021 recognition receptors (PPRs), including toll-like receptors (TLRs), NOD-like receptors (NLRs) and RIG-1-like receptors (RLRs), to screen for pathogen-associated molecular Publisher’s Note: MDPI stays neutral patterns (PAMPs) in the cytosol and extracellular matrix [5,6]. When cells detect a viral with regard to jurisdictional claims in pathogen, specific PRRs become activated that signal for mitochondrial antiviral signaling published maps and institutional affil- (MAVS) proteins to upregulate interferon-α/β (IFN-α/β) cytokine production [2,7–9]. iations. Cells then secrete IFN-α/β cytokines into the extracellular matrix to alert naive cells of an impending infection [10,11]. The production of IFN-α/β is one of the first lines of cellular defense against vi- ral pathogens. IFN-α/β activates the Janus kinase-signal transducer and activation of Copyright: © 2021 by the authors. transcription (JAK–STAT) signaling pathway (Figure1). When IFN- α/β binds to an IFN- Licensee MDPI, Basel, Switzerland. specific receptor on the cellular surface, a conformational change occurs that exposes sites This article is an open access article on the cytoplasmic part of the receptor that become phosphorylated by Janus kinase 1 distributed under the terms and (JAK1) and tyrosine-protein kinase 2 (TYK2). This phosphorylation attracts the nuclear conditions of the Creative Commons transcriptional regulators signal transducer and activator of transcription proteins 1 and 2 Attribution (CC BY) license (https:// (STAT1 and STAT2) to the cellular membrane where they are also phosphorylated by TYK2. creativecommons.org/licenses/by/ 4.0/). The phosphorylated STAT1 and STAT2 proteins form a ternary complex with methylated Viruses 2021, 13, 1102. https://doi.org/10.3390/v13061102 https://www.mdpi.com/journal/viruses Viruses 2021, 13, x FOR PEER REVIEW 2 of 19 Viruses 2021, 13, 1102 2 of 18 phosphorylated STAT1 and STAT2 proteins form a ternary complex with methylated in- terferoninterferon regulatory regulatory factor factor 9 9(IRF9), (IRF9), which which is is subsequently subsequently demethylated demethylated to to signal signal for for the migration of the STAT1–STAT2–IRF9 STAT1–STAT2–IRF9 (SSI)(SSI) complexcomplex toto thethe nucleus.nucleus. In the nucleus, the SSI complex binds to the ISG promoter region of the interferon-stimulated response element (ISRE), resulting in the transcriptional upregula upregulationtion of hundreds of ISGs that augment the antiviral immune response by halting ribosomal protein synthesis [[12,13],12,13], inducing the activation ofof regulatoryregulatory cell cell death death pathways pathways [14 [14–18],–18], and and carrying carrying out theout post-translationalthe post-transla- tionalmodification modification (PTM) (PTM) of host of and host viral and proteins viral proteins that impact that impact viral replication viral replication [19–25 ].[19–25]. Figure 1. Interferon-induced ISGylation of viral proteins. During the initial stages of infection, cellular IFN-α/β surface Figure 1. Interferon-induced ISGylation of viral proteins. During the initial stages of infection, cellular IFN-α/β surface receptors are bound by IFN-α/β-specific ligands that expose receptor phosphorylation sites to the cytoplasm. Janus kinase receptors are bound by IFN-α/β-specific ligands that expose receptor phosphorylation sites to the cytoplasm. Janus kinase 1 (JAK1) and tyrosine-protein kinase 2 (TYK2) phosphorylate exposed cytoplasmic IFN-α/β receptor sites, thus recruiting α β 1the (JAK1) nuclear and transcriptional tyrosine-protein regulators kinase2 sign (TYK2)al transducer phosphorylate and activator exposed of cytoplasmic transcription IFN- proteins/ receptor 1 and 2 sites,(STAT1 thus and recruiting STAT2) theto also nuclear be phosphorylated transcriptional regulatorsby TYK2. In signal the cytoplasm, transducer ph andosphorylated activator of transcriptionSTAT1 and STAT2 proteins form 1 and a ternary 2 (STAT1 complex and STAT2) with tomethylated also be phosphorylated IRF9 which is then by TYK2. demethylated In the cytoplasm, to signal fo phosphorylatedr the migration STAT1of the STAT1–STAT2–IRF9 and STAT2 form a ternary (SSI) complex complex to with the methylatednucleus. Upon IRF9 nuclear which entry, is then the demethylated SSI complex tobinds signal to forthe theISG migration promoter ofregion the STAT1–STAT2–IRF9 interferon-stimulated (SSI) response complex element to the nucleus.(ISRE) to Uponupregulate nuclear the entry, transcription the SSI complexof several binds hundred to the IS ISGGs, including promoter ISG15, region UBE1L, interferon-stimulated UBE2L6 and HERC5. response Translated element (ISRE)UBE1L, to UBE2L6 upregulate and theHERC5 transcription proteins ofare several subsequently hundred targeted ISGs, including to the perinuclear ISG15, UBE1L, regions UBE2L6 of the and cytoplasm HERC5. where Translated they UBE1L,work in UBE2L6tandem andto charge, HERC5 proteinstransfer areand subsequently attach ISG15 targeted onto respective to the perinuclear viral protei regionsn substrates of the cytoplasm to inhibit where viral they stability, work transport and reassembly. ME, methylation. This figure was created with Biorender™. in tandem to charge, transfer and attach ISG15 onto respective viral protein substrates to inhibit viral stability, transport and reassembly. ME, methylation. This figure was created with Biorender™. HECT and RCC1-containing protein 5 (HERC5) and interferon-stimulated gene 15 (ISG15)HECT are andtwo RCC1-containingantiviral immune protein proteins 5 (HERC5)that are induced and interferon-stimulated following IFN-α/β gene signal 15 transduction.(ISG15) are two Multiple antiviral studies immune have proteinsestablished that that are HERC5 induced plays following a central IFN- roleα/ inβ signalmam- maliantransduction. innate Multipleimmunity studies by ISGylating have established viral proteins that HERC5 to disrupt plays viral a central replication role in[26,27]. mam- Themalian antiviral innate function immunity of HERC5 by ISGylating was first viral discovered proteins toby disruptresearchers viral who replication found that [26,27 its]. co-expressionThe antiviral functionwith the ofE1 HERC5 ubiquitin-like was first modi discoveredfier activating by researchers enzyme UBE1L who found (Uba7; that E.C. its 6.2.1.45)co-expression and UBE2L6 with the (UbcH8; E1 ubiquitin-like E.C. 2.3.2.23) modifier resulted activating in normal enzyme rates of UBE1Lcellular (Uba7;ISGylation E.C. activity,6.2.1.45) despite and UBE2L6 the absence (UbcH8; of E.C.IFN- 2.3.2.23)α/β stimulation resulted [28]. in normal rates of cellular ISGylation activity,The despitefunctional the absencecharacterization of IFN-α of/β phosphokinasestimulation [28 signal]. transduction pathways has now Theclarified functional that UBE1L, characterization UBE2L6, HERC5 of phosphokinase and ISG15 signalexpression transduction are predominantly pathways hasin- ducednow clarified by the JAK–STAT that UBE1L, signaling UBE2L6, pathway HERC5 (Figure and ISG15 1) [29,30]. expression However, are in predominantly recent years, additionalinduced
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