Identification and Characterization of a Poliovirus Capsid Mutant with Enhanced Thermal Stability

Nguyen, Y; Jesudhasan, Palmy R.; Aguilera, Elizabeth R.; Pfeiffer, Julie K.

doi:10.1128/jvi.01510-18

Cited by 17 publications

(30 citation statements)

References 19 publications

(36 reference statements)

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“…3C and E). We also demonstrated that PV-M132V was stable during 8 days of incubation at 37°C in both PBS and feces (13). Interestingly, mengovirus exhibited significant inactivation after 4 days at 37°C, but incubation in feces limited this inactivation (Fig.…”

Section: Resultsmentioning

confidence: 77%

“…1B and Table 1). Additionally, our panel included a PV mutant with a single amino acid change in the VP1 capsid coding region (PV-M132V), which confers enhanced thermal stability in the absence of bacteria (13).…”

Section: Resultsmentioning

confidence: 99%

“…Interestingly, Aichi virus and mengovirus were very stable in PBS under these conditions. We also tested a recently identified heat-resistant PV mutant, PV-M132V (13). Like Aichi virus and mengovirus, PV-M132V was resistant to heat treatment, and thus, incubation with any of the compounds or bacterial strains did not increase stability (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Bacterial Stabilization of a Panel of Picornaviruses

Aguilera

Nguyen

Sasaki

et al. 2019

mSphere

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Several viruses encounter various bacterial species within the host and in the environment. Despite these close encounters, the effects of bacteria on picornaviruses are not completely understood. Previous work determined that poliovirus (PV), an enteric virus, has enhanced virion stability when exposed to bacteria or bacterial surface polysaccharides such as lipopolysaccharide. Virion stabilization by bacteria may be important for interhost transmission, since a mutant PV with reduced bacterial binding had a fecal-oral transmission defect in mice. Therefore, we investigated whether bacteria broadly enhance stability of picornaviruses from three different genera: Enterovirus (PV and coxsackievirus B3 [CVB3]), Kobuvirus (Aichi virus), and Cardiovirus (mengovirus). Furthermore, to delineate strain-specific effects, we examined two strains of CVB3 and a PV mutant with enhanced thermal stability. We determined that specific bacterial strains enhance thermal stability of PV and CVB3, while mengovirus and Aichi virus are stable at high temperatures in the absence of bacteria. Additionally, we determined that bacteria or lipopolysaccharide can stabilize PV, CVB3, Aichi virus, and mengovirus during exposure to bleach. These effects are likely mediated through direct interactions with bacteria, since viruses bound to bacteria in a pulldown assay. Overall, this work reveals shared and distinct effects of bacteria on a panel of picornaviruses. IMPORTANCE Recent studies have shown that bacteria promote infection and stabilization of poliovirus particles, but the breadth of these effects on other members of the Picornaviridae family is unknown. Here, we compared the effects of bacteria on four distinct members of the Picornaviridae family. We found that bacteria reduced inactivation of all of the viruses during bleach treatment, but not all viral strains were stabilized by bacteria during heat treatment. Overall, our data provide insight into how bacteria play differential roles in picornavirus stability.

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Section: Resultsmentioning

confidence: 77%

Section: Resultsmentioning

confidence: 99%

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Bacterial Stabilization of a Panel of Picornaviruses

Aguilera

Nguyen

Sasaki

et al. 2019

mSphere

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“…In such directed evolution experiments, virus populations are grown under conditions that favor either the emergence or further optimization of the desired phenotype. For example, this process has been used to obtain live attenuated vaccines for numerous viruses by repeated growth at suboptimal conditions 3 , improving in vivo models by serial infection of the desired host 4-9 , isolating RNA viruses with high-fidelity polymerases by growth under condition of increased mutational load 10,11 , selection of viruses with improved oncolytic properties [12][13][14] , or isolation of viruses and virus-like particles of increased stability [15][16][17] . To date, these studies have relied upon the natural mutational processes of RNA viruses to generate the diversity upon which selection is applied.…”

Section: Introductionmentioning

confidence: 99%

Increased RNA virus population diversity improves adaptability

Mattenberger

Vila-Nistal

Geller

2020

Preprint

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The replication machinery of most RNA viruses lacks proofreading mechanisms. As a result, RNA virus populations harbor a large amount of genetic diversity that confers them the ability to rapidly adapt to changes in their environment. In this work, we investigate whether further increasing the initial population diversity of a model RNA virus can improve adaptation to a single selection pressure, thermal inactivation. For this, we experimentally increased the diversity of coxsackievirus B3 (CVB3) populations across the capsid region. We then compared the ability of such high diversity CVB3 populations to achieve resistance to thermal inactivation relative to standard CVB3 populations in an experimental evolution setting. We find that high diversity viral populations are better able to achieve resistance to thermal inactivation at both the temperature employed during experimental evolution as well as at a more extreme temperature. Moreover, we identify mutations in the CVB3 capsid that confer resistance to thermal inactivation, finding significant mutational epistasis. Our results indicate that even naturally diverse RNA virus populations can benefit from experimental augmentation of population diversity for optimal adaptation and support the use of such viral populations in directed evolution efforts that aim to select for viruses with desired characteristics.

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“…It is reasonable to assume that the initial rotation, as well as the structural disruptions, may be inhibited by stronger interaction forces at the subunit interfaces. Experimental evolution experiments have shown that thermostable poliovirus variants carry mutations in the hydrophobic pocket region of VP1 (20)(21)(22). These mutations are distant from the pentameric interfaces but close to the interface of the protomeric subunits.…”

mentioning

confidence: 99%

Salt Enhances the Thermostability of Enteroviruses by Stabilizing Capsid Protein Interfaces

Meister

Prunotto

Peraro

et al. 2020

J Virol

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Enteroviruses are common agents of infectious disease that are spread by the fecal-oral route. They are readily inactivated by mild heat, which causes the viral capsid to disintegrate or undergo conformational change. While beneficial for the thermal treatment of food or water, this heat sensitivity poses challenges for the stability of enterovirus vaccines. The thermostability of an enterovirus can be modulated by the composition of the suspending matrix, though the effects of the matrix on virus stability are not understood. Here, we determined the thermostability of four enterovirus strains in solutions with various concentrations of NaCl and different pH values. The experimental findings were combined with molecular modeling of the protein interaction forces at the pentamer and the protomer interfaces of the viral capsids. While pH only had a modest effect on thermostability, increasing NaCl concentrations raised the breakpoint temperatures of all viruses tested by up to 20°C. This breakpoint shift could be explained by an enhancement of the van der Waals attraction forces at the two protein interfaces. In comparison, the (net repulsive) electrostatic interactions were less affected by NaCl. Depending on the interface considered, the breakpoint temperature shifted by 7.5 or 5.6°C per 100-kcal/(mol·Å) increase in protein interaction force. IMPORTANCE The genus Enterovirus encompasses important contaminants of water and food (e.g., coxsackieviruses), as well as viruses of acute public health concern (e.g., poliovirus). Depending on the properties of the surrounding matrix, enteroviruses exhibit different sensitivities to heat, which in turn influences their persistence in the environment, during food treatment, and during vaccine storage. Here, we determined the effect of NaCl and pH on the heat stability of different enteroviruses and related the observed effects to changes in protein interaction forces in the viral capsid. We demonstrate that NaCl renders enteroviruses thermotolerant and that this effect stems from an increase in van der Waals forces at different protein subunits in the viral capsid. This work sheds light on the mechanism by which salt enhances virus stability.

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Identification and Characterization of a Poliovirus Capsid Mutant with Enhanced Thermal Stability

Cited by 17 publications

References 19 publications

Bacterial Stabilization of a Panel of Picornaviruses

Bacterial Stabilization of a Panel of Picornaviruses

Increased RNA virus population diversity improves adaptability

Salt Enhances the Thermostability of Enteroviruses by Stabilizing Capsid Protein Interfaces

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