A Viral RNA Structural Element Alters Host Recognition of Nonself RNA

Hyde, Jennifer; Gardner, Christina L.; Kimura, Taishi; White, James P.; Liu, Gai; Trobaugh, Derek W.; Huang, Cheng; Tonelli, Marco; Paessler, Slobodan; Takeda, Kiyoshi; Klimstra, William B.; Amarasinghe, Gaya K.; Diamond, Michael S.

doi:10.1126/science.1248465

Cited by 156 publications

(200 citation statements)

References 46 publications

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“…To extend these findings to another family of RNA viruses that cause CNS disease, we evaluated the pathogenesis of VEEV, an encephalitic alphavirus, in Asyn-knockout mice. We used an attenuated strain of VEEV (TC83) that cannot escape IFIT1 restriction (33). Using VEEV TC83, we found that the virus grew to titers of up to 10 8 PFU per gram of brain tissue in the brains of Asyn-knockout mice, and Asyn-knockout mice experienced significant disease from VEEV TC83, as evidenced by increased weight loss compared to that found in TC83-inoculated Snca Ϫ/ϩ mice.…”

Section: Discussionmentioning

confidence: 99%

“…Using a neuroinvasive WNV strain, we found that Asyn expression inhibited the growth of infectious viral particles in the brain. We next used a vaccine strain of an alphavirus, Venezuelan equine encephalitis virus (VEEV; strain TC83), to determine if Asyn expression inhibits the growth of virus from a different enveloped RNA virus family not related to WNV (33 ) from TC83-infected Snca Ϫ/Ϫ mice was significantly (P ϭ 0.03, ANOVA with the Mann-Whitney test) increased compared with that in striatal and cortical tissue from Snca Ϫ/ϩ mice, in which virus was undetectable (Fig. 5A).…”

Section: Resultsmentioning

confidence: 99%

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Alpha-Synuclein Expression Restricts RNA Viral Infections in the Brain

Beatman

Massey

Shives

et al. 2016

J Virol

187

176

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We have discovered that native, neuronal expression of alpha-synuclein (Asyn) inhibits viral infection, injury, and disease in the central nervous system (CNS). Enveloped RNA viruses, such as West Nile virus (WNV), invade the CNS and cause encephalitis, yet little is known about the innate neuron-specific inhibitors of viral infections in the CNS. Following WNV infection of primary neurons, we found that Asyn protein expression is increased. The infectious titer of WNV and Venezuelan equine encephalitis virus (VEEV) TC83 in the brains of Asyn-knockout mice exhibited a mean increase of 10 4.5 infectious viral particles compared to the titers in wild-type and heterozygote littermates. Asyn-knockout mice also exhibited significantly increased virusinduced mortality compared to Asyn heterozygote or homozygote control mice. Virus-induced Asyn localized to perinuclear, neuronal regions expressing viral envelope protein and the endoplasmic reticulum (ER)-associated trafficking protein Rab1. In Asyn-knockout primary neuronal cultures, the levels of expression of ER signaling pathways, known to support WNV replication, were significantly elevated before and during viral infection compared to those in Asyn-expressing primary neuronal cultures. We propose a model in which virus-induced Asyn localizes to ER-derived membranes, modulates virus-induced ER stress signaling, and inhibits viral replication, growth, and injury in the CNS. These data provide a novel and important functional role for the expression of native alpha-synuclein, a protein that is closely associated with the development of Parkinson's disease. IMPORTANCENeuroinvasive viruses such as West Nile virus are able to infect neurons and cause severe disease, such as encephalitis, or infection of brain tissue. Following viral infection in the central nervous system, only select neurons are infected, implying that neurons exhibit innate resistance to viral infections. We discovered that native neuronal expression of alpha-synuclein inhibited viral infection in the central nervous system. When the gene for alpha-synuclein was deleted, mice exhibited significantly decreased survival, markedly increased viral growth in the brain, and evidence of increased neuron injury. Virus-induced alphasynuclein localized to intracellular neuron membranes, and in the absence of alpha-synuclein expression, specific endoplasmic reticulum stress signaling events were significantly increased. We describe a new neuron-specific inhibitor of viral infections in the central nervous system. Given the importance of alpha-synuclein as a cause of Parkinson's disease, these data also ascribe a novel functional role for the native expression of alpha-synuclein in the CNS.

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Section: Discussionmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Alpha-Synuclein Expression Restricts RNA Viral Infections in the Brain

Beatman

Massey

Shives

et al. 2016

J Virol

187

176

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“…Type I interferon secretion leads to the transcription of hundreds of interferon-stimulated genes, which display antiviral properties by targeting almost any step of the virus replication cycle (66). Interestingly, Hyde and colleagues demonstrated that alphaviruses, which are also single-stranded, positive-strand RNA viruses, use mutations within the 5= noncoding region affecting secondary-structural elements of their RNAs to alter interferon-stimulated protein binding and functions (67). Their results suggest an evasion mechanism by which a deleted virus with modified 5= RNA secondary structures could avoid immune restriction, despite type 1 IFN secretion and interferon-stimulated gene transcription, leading to long-term virus persistence in the heart.…”

Section: Discussionmentioning

confidence: 99%

Functional Consequences of RNA 5′-Terminal Deletions on Coxsackievirus B3 RNA Replication and Ribonucleoprotein Complex Formation

Lévêque

Garcia

Bouin

et al. 2017

J Virol

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Group B coxsackieviruses are responsible for chronic cardiac infections. However, the molecular mechanisms by which the virus can persist in the human heart long after the signs of acute myocarditis have abated are still not completely understood. Recently, coxsackievirus B3 strains with 5=-terminal deletions in genomic RNAs were isolated from a patient suffering from idiopathic dilated cardiomyopathy, suggesting that such mutant viruses may be the forms responsible for persistent infection. These deletions lacked portions of 5= stem-loop I, which is an RNA secondary structure required for viral RNA replication. In this study, we assessed the consequences of the genomic deletions observed in vivo for coxsackievirus B3 biology. Using cell extracts from HeLa cells, as well as transfection of luciferase replicons in two types of cardiomyocytes, we demonstrated that coxsackievirus RNAs harboring 5= deletions ranging from 7 to 49 nucleotides in length can be translated nearly as efficiently as those of wild-type virus. However, these 5= deletions greatly reduced the synthesis of viral RNA in vitro, which was detected only for the 7-and 21-nucleotide deletions. Since 5= stem-loop I RNA forms a ribonucleoprotein complex with cellular and viral proteins involved in viral RNA replication, we investigated the binding of the host cell protein PCBP2, as well as viral protein 3CD pro , to deleted positive-strand RNAs corresponding to the 5= end. We found that binding of these proteins was conserved but that ribonucleoprotein complex formation required higher PCBP2 and 3CD pro concentrations, depending on the size of the deletion. Overall, this study confirmed the characteristics of persistent CVB3 infection observed in heart tissues and provided a possible explanation for the low level of RNA replication observed for the 5=-deleted viral genomes-a less stable ribonucleoprotein complex formed with proteins involved in viral RNA replication.IMPORTANCE Dilated cardiomyopathy is the most common indication for heart transplantation worldwide, and coxsackie B viruses are detected in about one-third of idiopathic dilated cardiomyopathies. Terminal deletions at the 5= end of the viral genome involving an RNA secondary structure required for RNA replication have been recently reported as a possible mechanism of virus persistence in the human heart. These mutations are likely to disrupt the correct folding of an RNA secondary structure required for viral RNA replication. In this report, we demonstrate that transfected RNAs harboring 5=-terminal sequence deletions are able to direct the synthesis of viral proteins, but not genomic RNAs, in human and murine cardiomyocytes. Moreover, we show that the binding of cellular and viral replication factors to viral RNA is conserved despite genomic deletions but that the impaired RNA synthesis as-

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“…Chimeric flaviviruses have been engineered to study pathogenesis (Yoshii et al, 2014), replication regulation and virushost interaction (Courtney et al, 2012;Evans & Seeger, 2007;Hyde et al, 2014). Especially, using live attenuated strains as the genetic backbone, multiple versions of chimeric flaviviruses have been successfully designed and well-explored in the development of vaccines against DENV, WNV, JEV and TBEV (Brandler et al, 2005;Chambers et al, 1999;Guirakhoo et al, 2000;Guy & Jackson, 2016;Pletnev et al, 2002;Wright et al, 2008).…”

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confidence: 99%