Functional role of the 5′ terminal cloverleaf in Coxsackievirus RNA replication

Sharma, Nidhi; Ogram, Sushma A.; Morasco, Benjamin J.; Spear, Allyn; Chapman, Nora M.; Flanegan, James B.

doi:10.1016/j.virol.2009.07.039

Cited by 38 publications

(61 citation statements)

References 41 publications

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“…Other studies have detected very low levels of replication of TD RNAs in HeLa S10 cell in vitro RNA synthesis reactions or with semi-nested reverse transcription (RT)-PCR amplification, even for the deletion of 49 nucleotides (32,34,48). The most likely explanation for these differences is that the methods used in our study were not sufficiently sensitive to detect the very low levels of RNA replication directed by the TD30 and TD49 RNAs in vitro.…”

Section: Discussionmentioning

confidence: 56%

“…Deletions involving sequences within the 5= S-L I structure of genomic RNAs are known to cause profound changes in viral biology by reducing viral replication compared to wild-type viruses (3,(32)(33)(34)(35). Indeed, the discovery of TD mutations in heart biopsy specimens was associated with low viral loads and abnormal positive-strand/ negative-strand viral RNA ratios, close to 1 rather than the high ratios (30 to 70) normally observed in enterovirus-infected cells with wild-type viruses (32,(36)(37)(38).…”

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confidence: 99%

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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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Section: Discussionmentioning

confidence: 56%

mentioning

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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“…However, host survival was commonly observed in parallel with detectable levels of viral RNA in the adult CNS for at least 90 days postinfection (p.i.). The ability of CVB3 to persist in other organs, in particular, the heart, has been well documented (6,25) and may involve genetic alterations in the virus which may limit replication and cytopathic effects (CPE) in the host cell (23,24,36). We hypothesized that the continued presence of viral RNA and/or viral gene products may affect normal neural stem cell migration and/or differentiation in the developing CNS.…”

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confidence: 99%

Coxsackievirus Preferentially Replicates and Induces Cytopathic Effects in Undifferentiated Neural Progenitor Cells

Tsueng

Tabor-Godwin

Gopal

et al. 2011

J Virol

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Enteroviruses, including coxsackieviruses, exhibit significant tropism for the central nervous system, and these viruses are commonly associated with viral meningitis and encephalitis. Previously, we described the ability of coxsackievirus B3 (CVB3) to infect proliferating neuronal progenitor cells located in the neonatal subventricular zone and persist in the adult murine central nervous system (CNS). Here, we demonstrate that cultured murine neurospheres, which comprise neural stem cells and their progeny at different stages of development, were highly susceptible to CVB3 infection. Neurospheres, or neural progenitor and stem cells (NPSCs), isolated from neonatal C57BL/6 mice, supported high levels of infectious virus production and high viral protein expression levels following infection with a recombinant CVB3 expressing enhanced green fluorescent protein (

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“…2). Mutagenesis studies on PV and CVB-3 have demonstrated that the presence of the 5NCL is crucial for the synthesis of both negative and positive-strand RNAs and VPg uridylylation (Sharma et al, 2009;Vogt and Andino, 2010;Ogram and Flanegan, 2011). The cloverleaf structure is composed of four stem-loop elements (SL a-d).…”

Section: Structural Elements Of Rna Involved In Viral Replicationmentioning

confidence: 99%

Molecular mechanisms of genome expression of coxsackievirus B3 that belongs to enteroviruses

Dutkiewicz¹,

Świątkowska²,

Ojdowska³

et al. 2012

bta

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Coxsackievirus B3 (CVB-3) belongs to the Picornaviridae family of enterovirus genus of pathogens that cause a great number of human diseases. A viral infection is associated with many pathological states such as: myocarditis, dilated cardiomyopathy, pericarditis, pleurodynia, systemic infection in infants, aseptic meningitis, and pancreatitis. Since viral diseases, especially in their chronic state, are difficult to treat, there has not been as yet, any specific therapeutic developed against coxsackievirus till date. CVB-3 is a single stranded, positive-sense RNA virus that encodes one large open reading frame flanked by two untranslated regions (UTR). The 5NUTR contains an IRES element that directs the translation process and a cloverleaf structure that regulate viral replication. The complementary, 3N terminal region of the replicative strand is also believed to be crucial for the replication of events. The secondary structure RNA elements regulate the most important processes in the viral propagation cycle. The mechanisms that rule the CBV-3 gene expression, its genome structure and the key steps of its viral life cycle are being reviewed in the hope that better knowledge of these processes will lead to better understanding of the molecular biology of CVB-3 and to the design of an effective therapy against this enterovirus.

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Functional role of the 5′ terminal cloverleaf in Coxsackievirus RNA replication

Cited by 38 publications

References 41 publications

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

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

Coxsackievirus Preferentially Replicates and Induces Cytopathic Effects in Undifferentiated Neural Progenitor Cells

Molecular mechanisms of genome expression of coxsackievirus B3 that belongs to enteroviruses

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