Poliovirus 2C region functions during encapsidation of viral RNA

Vance, L M; Moscufo, Nicola; Chow, Marie; Heinz, Beverly A.

doi:10.1128/jvi.71.11.8759-8765.1997

Cited by 106 publications

(63 citation statements)

References 37 publications

(61 reference statements)

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“…2C ATPase is a complex nonstructural polypeptide that expresses numerous functions: (i) it has been proven to be crucial for RNA replication and binding (120-124); (ii) it contains a nucleoside triphosphate (NTP)-binding domain (125) and possesses ATPase activity (discovered in vitro), which is inhibited by guanidine hydrochloride (GnHCl) (126); (iii) it has been implicated (weakly) in virus uncoating (127); (iv) together with its processing precursor 2BC ATPase , it is involved in cellular membrane rearrangements and the formation of the cytoplasmic vesicle "web" typical of all enterovirus replication (13,(128)(129)(130)(131); and (v) drug (hydantoin) inhibition and genetic studies have provided convincing genetic evidence that 2C ATPase is involved in encapsidation (20,21,132).…”

Section: Multiple Roles Of Enterovirus 2c Atpase In the Virus Replicamentioning

confidence: 99%

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Picornavirus Morphogenesis

Jiang

Liu

et al. 2014

Microbiol Mol Biol Rev

184

196

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SUMMARY The Picornaviridae represent a large family of small plus-strand RNA viruses that cause a bewildering array of important human and animal diseases. Morphogenesis is the least-understood step in the life cycle of these viruses, and this process is difficult to study because encapsidation is tightly coupled to genome translation and RNA replication. Although the basic steps of assembly have been known for some time, very few details are available about the mechanism and factors that regulate this process. Most of the information available has been derived from studies of enteroviruses, in particular poliovirus, where recent evidence has shown that, surprisingly, the specificity of encapsidation is governed by a viral protein-protein interaction that does not involve an RNA packaging signal. In this review, we make an attempt to summarize what is currently known about the following topics: (i) encapsidation intermediates, (ii) the specificity of encapsidation (iii), viral and cellular factors that are required for encapsidation, (iv) inhibitors of encapsidation, and (v) a model of enterovirus encapsidation. Finally, we compare some features of picornavirus morphogenesis with those of other plus-strand RNA viruses.

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Section: Multiple Roles Of Enterovirus 2c Atpase In the Virus Replicamentioning

confidence: 99%

“…The small organic compound hydantoin [5(3,4-dichlorophenyl) methylhydantoin] strongly inhibits enterovirus morphogenesis in tissue culture by aborting maturation at an 110S intermediate (132). The effect of hydantoin is reversible since the 110S particles can be chased into mature virions after removal of the drug.…”

Section: Hydantoinmentioning

confidence: 99%

Picornavirus Morphogenesis

Jiang

Liu

et al. 2014

Microbiol Mol Biol Rev

184

196

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“…To test if the growth defect we observed in the absence of TDP2 was due to premature packaging of the viral RNA, we used a known picornavirus encapsidation inhibitor called hydantoin [60], which at low concentrations (<50 µg/mL) blocks virion maturation but leaves both virus translation and RNA synthesis unaffected. At concentrations >50 µg/mL, however, RNA synthesis is also diminished [61].…”

Section: Premature Encapsidation Of Viral Rna Is Not Responsible For mentioning

confidence: 99%

Effects of TDP2/VPg Unlinkase Activity on Picornavirus Infections Downstream of Virus Translation

Holmes

Zagnoli-Vieira

Caldecott

et al. 2020

Viruses

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In this study, we characterized the role of host cell protein tyrosyl-DNA phosphodiesterase 2 (TDP2) activity, also known as VPg unlinkase, in picornavirus infections in a human cell model of infection. TDP2/VPg unlinkase is used by picornaviruses to remove the small polypeptide, VPg (Virus Protein genome-linked, the primer for viral RNA synthesis), from virus genomic RNA. We utilized a CRISPR/Cas-9-generated TDP2 knock out (KO) human retinal pigment epithelial-1 (hRPE-1) cell line, in addition to the wild type (WT) counterpart for our studies. We determined that in the absence of TDP2, virus growth kinetics for two enteroviruses (poliovirus and coxsackievirus B3) were delayed by about 2 h. Virus titers were reduced by ~2 log10 units for poliovirus and 0.5 log10 units for coxsackievirus at 4 hours post-infection (hpi), and by ~1 log10 unit at 6 hpi for poliovirus. However, virus titers were nearly indistinguishable from those of control cells by the end of the infectious cycle. We determined that this was not the result of an alternative source of VPg unlinkase activity being activated in the absence of TPD2 at late times of infection. Viral protein production in TDP2 KO cells was also substantially reduced at 4 hpi for poliovirus infection, consistent with the observed growth kinetics delay, but reached normal levels by 6 hpi. Interestingly, this result differs somewhat from what has been reported previously for the TDP2 KO mouse cell model, suggesting that either cell type or species-specific differences might be playing a role in the observed phenotype. We also determined that catalytically inactive TDP2 does not rescue the growth defect, confirming that TDP2 5′ phosphodiesterase activity is required for efficient virus replication. Importantly, we show for the first time that polysomes can assemble efficiently on VPg-linked RNA after the initial round of translation in a cell culture model, but both positive and negative strand RNA production is impaired in the absence of TDP2 at mid-times of infection, indicating that the presence of VPg on the viral RNA affects a step in the replication cycle downstream of translation (e.g., RNA synthesis). In agreement with this conclusion, we found that double-stranded RNA production (a marker of viral RNA synthesis) is delayed in TDP2 KO RPE-1 cells. Moreover, we show that premature encapsidation of nascent, VPg-linked RNA is not responsible for the observed virus growth defect. Our studies provide the first lines of evidence to suggest that either negative- or positive-strand RNA synthesis (or both) is a likely candidate for the step that requires the removal of VPg from the RNA for an enterovirus infection to proceed efficiently.

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“…Mutation T173M in 2C conferred resistance to SMSK with an EC50 of 5.7±0.3 µM (~4-fold change over WT) when reversed-engineered into the WT virus backbone. Viruses resistant to 2C inhibitors have been reported for several compounds (TBZE-029, HBB, Guanidine HCl, hydantoin, fluoxetine) [26][27][28][34][35][36] . Some of these resistant viruses have an attenuated virus growth 28 or even a compound-dependent phenotype 27,37 .…”

Section: Identification Of Mutations In Double-resistant Populationsmentioning

confidence: 99%

Assessing the in vitro resistance development in Enterovirus 71 in the context of combination antiviral treatment

Lanko

Shi

Patil

et al. 2020

Preprint

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There are currently no antivirals available to treat infection with enterovirus A71 (EV-A71) or any other enterovirus. The extensively studied capsid binders select rapidly for drug-resistant variants. We here explore whether the combination of two direct-acting enterovirus inhibitors with a different mechanism of action may delay or prevent resistance development to the capsid binders. To that end, the in vitro dynamics of resistance development to the capsid binder pirodavir was studied either alone or in combination with (i) a viral 2C-targeting compound (SMSK_0213), (ii) a viral 3C-protease inhibitor (rupintrivir) or (iii) a viral RNAdependent RNA polymerase (RdRp) inhibitor [7-deaza-2'C-methyladenosine (7DMA)]. We demonstrate that combining pirodavir with either rupintrivir or the nucleoside analogue 7DMA delays the development of resistance to pirodavir and that no resistance to the protease or polymerase inhibitor develops. The combination of pirodavir with the 2C inhibitor results in a double-resistant virus population. The deep sequencing analysis of resistant populations revealed that even though resistant mutations are present in less than 30% of the population, this still provides the resistant phenotype.

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Poliovirus 2C region functions during encapsidation of viral RNA

Cited by 106 publications

References 37 publications

Picornavirus Morphogenesis

Picornavirus Morphogenesis

Effects of TDP2/VPg Unlinkase Activity on Picornavirus Infections Downstream of Virus Translation

Assessing the in vitro resistance development in Enterovirus 71 in the context of combination antiviral treatment

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