cGAS-STING Signaling Regulates Initial Innate Control of Cytomegalovirus Infection

Lio, Chan-Wang Jerry; McDonald, Bryan; Takahashi, Mariko; Dhanwani, Rekha; Sharma, Nikita; Huang, Junhua; Pham, Elise; Benedict, Chris A.; Sharma, Sonia

doi:10.1128/jvi.01040-16

Cited by 111 publications

(122 citation statements)

References 47 publications

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“…With this in mind, host signaling pathways necessary for HCMV-induced ISG15 accumulation and conjugation were examined. For HCMV, engaging the cytoplasmic dsDNA sensor cGAS is responsible for activating the critical adaptor protein STING and triggering antiviral responses (34,35). To determine if ISG15 accumulation was cGAS dependent, NHDFs transfected with control NS siRNA or cGAS-specific siRNAs were infected with HCMV, and ISG15 abundance was measured by immunoblotting.…”

Section: Resultsmentioning

confidence: 99%

Restriction of Human Cytomegalovirus Replication by ISG15, a Host Effector Regulated by cGAS-STING Double-Stranded-DNA Sensing

Bianco

Mohr

2017

J Virol

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Accumulation of the interferon-stimulated gene 15 (ISG15) protein product, which is reversibly conjugated to numerous polypeptide targets, impacts the proteome and physiology of uninfected and infected cells. While many viruses, including human cytomegalovirus (HCMV), blunt host antiviral defenses by limiting ISG expression, the overall abundance of ISG15 monomer and protein conjugates rises in HCMV-infected cells. However, the molecular signals underlying ISG15 accumulation and whether the ISG15 polypeptide itself influences HCMV infection biology remain unknown. Here, we establish that the ISG15 gene product itself directly regulates HCMV replication and that its accumulation restricts productive virus growth. Although ISG15 monomer and protein conjugate accumulation was induced in cells infected with UV-inactivated HCMV, it was subsequently reduced, but not eliminated, by an immediate-early (IE) or early (E) virus-encoded function(s). Instead, HCMV-induced ISG15 monomer and protein conjugate accumulation was dependent upon the double-stranded DNA (dsDNA) sensor cyclic GMP-AMP synthase (cGAS), the innate immune adaptor STING, and interferon signaling. Significantly, dsDNA itself was sufficient to induce cGAS-, STING-, and interferon signaling-dependent ISG15 monomer and conjugate protein accumulation in uninfected cells. Accumulation of ISGylated proteins in uninfected cells treated with dsDNA was prevented by expressing the HCMV multifunctional IE1 transactivator. This demonstrates that expression of a single host interferon-stimulated gene, ISG15, restricts HCMV replication, and that IE1 is sufficient to blunt ISGylation in response to dsDNA sensing in uninfected cells. Moreover, it establishes that ISGylation modifies the proteomes of virus-infected and uninfected normal cells in response to cell-intrinsic dsDNA sensing dependent upon cGAS-STING.IMPORTANCE By antagonizing type I interferon production and action, many viruses, including human cytomegalovirus (HCMV), evade host defenses. However, levels of the interferon-induced ISG15 protein, which is covalently conjugated to host and viral proteins, increase in HCMV-infected cells. How ISG15 accumulation is regulated and whether the ISG15 polypeptide influences HCMV replication remain unknown. This study establishes that ISG15 itself restricts HCMV replication and that HCMV-induced ISG15 accumulation is triggered by host defenses that detect cytoplasmic double-stranded DNA (dsDNA). Remarkably, dsDNA triggered ISG15 accumulation even in uninfected cells, and this was reduced by HCMV IE1 expression. This shows that ISG15 itself controls the replication of HCMV, which causes life-threatening disease among the immunocompromised and is a significant source of congenital morbidity and mortality among newborns. Moreover, it demonstrates that ISG15 modifies the uninfected cell proteome in response to dsDNA, potentially impacting responses to DNA vaccines, gene therapy, and autoimmune disease pathogenesis.

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Section: Resultsmentioning

confidence: 99%

Restriction of Human Cytomegalovirus Replication by ISG15, a Host Effector Regulated by cGAS-STING Double-Stranded-DNA Sensing

Bianco

Mohr

2017

J Virol

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“…NOD2-deficient mice had enhanced susceptibility to infection with respiratory syncytial virus (RSV), decreased IRF3 phosphorylation, and type I IFN production. Redundancy of innate immune response pathways to herpesviruses is well known, and some of the recently described pattern recognition receptors, such as IFI16 and cGMP-AMP synthase (cGAS), appear to be broad sensors of different herpesviruses (29,(36)(37)(38)(39)(40)(41). In the case of NOD1, specific HCMV suppression through NOD1 activation (but not HSV-1 suppression) suggests the possible use of specialized pathways through HCMV which could be targeted for virus control.…”

Section: Discussionmentioning

confidence: 99%

Role of nucleotide-binding oligomerization domain 1 (NOD1) and its variants in human cytomegalovirus control in vitro and in vivo

Fan

Roy

Mukhopadhyay

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Induction of nucleotide-binding oligomerization domain 2 (NOD2) and downstream receptor-interacting serine/threonine-protein kinase 2 (RIPK2) by human cytomegalovirus (HCMV) is known to up-regulate antiviral responses and suppress virus replication. We investigated the role of nucleotide-binding oligomerization domain 1 (NOD1), which also signals through RIPK2, in HCMV control. NOD1 activation by Tri-DAP (NOD1 agonist) suppressed HCMV and induced IFN-β. Mouse CMV was also inhibited through NOD1 activation. NOD1 knockdown (KD) or inhibition of its activity with small molecule ML130 enhanced HCMV replication in vitro. NOD1 mutations displayed differential effects on HCMV replication and antiviral responses. In cells overexpressing the E56K mutation in the caspase activation and recruitment domain, virus replication was enhanced, but in cells overexpressing the E266K mutation in the nucleotidebinding domain or the wild-type NOD1, HCMV was inhibited, changes that correlated with IFN-β expression. The interaction of NOD1 and RIPK2 determined the outcome of virus replication, as evidenced by enhanced virus growth in NOD1 E56K mutant cells (which failed to interact with RIPK2). NOD1 activities were executed through IFN-β, given that IFN-β KD reduced the inhibitory effect of Tri-DAP on HCMV. Signaling through NOD1 resulting in HCMV suppression was IKKα-dependent and correlated with nuclear translocation and phosphorylation of IRF3. Finally, NOD1 polymorphisms were significantly associated with the risk of HCMV infection in women who were infected with HCMV during participation in a glycoprotein B vaccine trial. Collectively, our data indicate a role for NOD1 in HCMV control via RIPK2-IKKα-IRF3 and suggest that its polymorphisms predict the risk of infection.H uman cytomegalovirus (HCMV), a member of the herpesvirus family, induces complex innate immune responses (1, 2). Despite this effective and multifaceted induction, HCMV has developed strategies to counteract its recognition (3), allowing for its productive replication and the establishment of latency. Identification and characterization of HCMV-induced innate immune responses and resulting signaling pathways may provide novel strategies for its control.Mounting evidence indicates that HCMV sensing is an intricate process involving activities of membrane, cytoplasmic, and nuclear receptors. Several HCMV-encoded proteins directly activate innate immune response molecules; the glycoprotein B (gB) binds to and activates TLR2 (4), and pp65 interacts with IFI16 (5). Other viral proteins, dsDNA, or dsRNA are likely to activate or inhibit host innate response molecules. Several previous reports have highlighted a complex role of the IFN pathway in response to HCMV. The activity of the promyeolcytic leukemia protein, a regulator of type I IFN response, is counteracted by HCMV-encoded immediate early 1 protein (IE1) (6). A cytoplasmic dsDNA sensor, ZBP1, activates IRF3 on infection, and its overexpression inhibits HCMV replication (7). IFN-inducible protein IFI16 modestly in...

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“…In mice, studies have shown a direct interaction between murine cytomegalovirus (MCMV) and NK cells, where the activating NK cell receptor Ly49H recognizes the m157 glycoprotein, thereby activating the NK cell to produce cytokines important for antiviral immunity and/or to kill (Krug et al 2004;Varani et al 2007;Zucchini et al 2008) MCMV and HCM-V AIM2 dsDNA (Rathinam et al 2010;Huang et al 2017) MCMV and HCM-V cGAS-STI-NG dsDNA (Lio et al 2016) MCMV TLR3 dsRNA (Tabeta et al 2004) MCMV TLR7 ssRNA (Zucchini et al 2008) HCMV DAI dsDNA (DeFilippis et al 2010) HCMV IFI16 dsDNA (Gariano et al 2012) the virus-infected cell through release of perforins and granzymes (Brown et al 2001;Pyzik et al 2011). Indeed, m157 deletion mutant MCMV fails to activate Ly49H+ NK cells and results in enhanced virulence (Voigt et al 2003;Bubić et al 2004).…”

Section: Innate Immunitymentioning

confidence: 99%

A constant companion: immune recognition and response to cytomegalovirus with aging and implications for immune fitness

Souquette¹,

Frere²,

Smithey³

et al. 2017

GeroScience

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Approximately 50% of individuals aged 6-49 years in the United States are infected with cytomegalovirus (CMV), with seroprevalence increasing with age, reaching 85-90% by 75-80 years according to Bate et al. (Clin Infect Dis 50 (11): 1439-1447 and Pawelec et al. (Curr Opin Immunol 24:507-511, 2012). Following primary infection, CMV establishes lifelong latency with periodic reactivation. Immunocompetent hosts experience largely asymptomatic infection, but CMV can cause serious illness in immunocompromised populations, such as transplant patients and the elderly. Control of CMV requires constant immune surveillance, and recent discoveries suggest this demand alters general features of the immune system in infected individuals. Here, we review recent advances in the understanding of the immune response to CMV and the role of CMV in immune aging and fitness, while highlighting the importance of potential confounding factors that influence CMV studies. Understanding how CMV contributes to shaping Bbaseline^immunity has important implications for a host's ability to mount effective responses to diverse infections and vaccination.

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cGAS-STING Signaling Regulates Initial Innate Control of Cytomegalovirus Infection

Cited by 111 publications

References 47 publications

Restriction of Human Cytomegalovirus Replication by ISG15, a Host Effector Regulated by cGAS-STING Double-Stranded-DNA Sensing

Restriction of Human Cytomegalovirus Replication by ISG15, a Host Effector Regulated by cGAS-STING Double-Stranded-DNA Sensing

Role of nucleotide-binding oligomerization domain 1 (NOD1) and its variants in human cytomegalovirus control in vitro and in vivo

A constant companion: immune recognition and response to cytomegalovirus with aging and implications for immune fitness

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