An increased replication fidelity mutant of foot-and-mouth disease virus retains fitness in vitro and virulence in vivo

Zeng, Jianxiong; Wang, Haiwei; Xie, Xing; Yang, Decheng; Zhou, Guohui; Yu, Li

doi:10.1016/j.antiviral.2013.07.008

Cited by 42 publications

(48 citation statements)

References 36 publications

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“…Although the A38V mutant polymerase had increased replication fidelity, it was not resistant to 50 M ribavirin, contradicting the hypothesis that enhanced polymerase fidelity confers resistance to mutagens (23,24,31). This led us to investigate the underlying mechanism of this unexpected phenomenon.…”

Section: Generation Of Fmdv Populations Resistant To Ribavirinmentioning

confidence: 71%

“…High-fidelity FMDV RdRp mutants isolated by mutagen selection have not been reported (25,30). We recently isolated the FMDV high-fidelity R84H mutant using 5-FU, but unfortunately, this mutant did not exhibit an attenuated phenotype, possibly due to a moderate increase in replication fidelity (31). Obviously, these observations are not enough to clearly show the correlation between the increase in replication fidelity or the restriction in quasispecies diversity and viral phenotypes, including fitness and virulence.…”

mentioning

confidence: 62%

“…These variants are ideal tools to examine viral evolutionary dynamics. Studies of high-fidelity RdRp mutants of PV, CVB3, CHIKV, and FMDV have demonstrated that (i) increased polymerase fidelity results in viral tolerance to mutagens, (ii) different yet broad-spectrum mutagen resistance occurs, and (iii) not all mutants are attenuated in animal models (23,24,27,28,31). In this study, attenuated FMDV high-fidelity RdRp variants showing different degrees of polymerase fidelity were isolated and examined in the context of polymerase fidelity and mutagen resistance.…”

Section: Discussionmentioning

confidence: 99%

“…The FMDV R84H variant, which has a 1.44-fold increase in RdRp fidelity, is not attenuated in suckling mice (31). Compared with WT FMDV, the newly isolated RdRp D5N and A38V variants and the constructed DAMM variant showed 1.51-, 1.65-, and 1.88-fold enhancements in replication fidelity, respectively, and exhibited 10-, 20-, and 100-fold viral attenuation in suckling mice, respectively (Fig.…”

Section: Generation Of Fmdv Populations Resistant To Ribavirinmentioning

confidence: 99%

“…Previous studies of selected high-fidelity RdRp variants have consistently revealed that resistance to a mutagen(s) is associated with increased polymerase fidelity (23,24,27,31). To examine the replication fidelity of our RdRp mutants, their mutation frequency was determined in the absence or presence of ribavirin.…”

Section: Generation Of Fmdv Populations Resistant To Ribavirinmentioning

confidence: 99%

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Ribavirin-Resistant Variants of Foot-and-Mouth Disease Virus: the Effect of Restricted Quasispecies Diversity on Viral Virulence

Zeng

Wang

Xie

et al. 2014

J Virol

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Mutagenic nucleoside analogues can be used to isolate RNA virus high-fidelity RNA-dependent RNA polymerase (RdRp) variants, the majority of which are attenuated in vivo. However, attenuated foot-and-mouth disease virus (FMDV) high-fidelity RdRp variants have not been isolated, and the correlations between RdRp fidelity and virulence remain unclear. Here, the mutagen ribavirin was used to select a ribavirin-resistant population of FMDV, and 4 amino acid substitutions (D5N, A38V, M194I, and M296V) were identified in the RdRp-coding region of the population. Through single or combined mutagenesis using a reverse genetics system, we generated direct experimental evidence that the rescued D5N, A38V, and DAMM mutants but not the M194I and M296V mutants are high-fidelity RdRp variants. Mutagen resistance assays revealed that the higher replication fidelity was associated with higher-level resistance to ribavirin. In addition, significantly attenuated fitness and virulence phenotypes were observed for the D5N, A38V, and DAMM mutants. Based on a systematic quantitative analysis of fidelity and virulence, we concluded that higher replication fidelity is associated with a more attenuated virus. These data suggest that the resulting restricted quasispecies diversity compromises the adaptability and virulence of an RNA virus population. The modulation of replication fidelity to attenuate virulence may represent a general strategy for the rational design of new types of live, attenuated vaccine strains. IMPORTANCEThe ribavirin-isolated poliovirus (PV) RdRp G64S variant, the polymerases of which were of high replication fidelity, was attenuated in vivo. It has been proposed (M. Vignuzzi, E. Wendt, and R. Andino, Nat. Med. 14:154 -161, http://dx.doi.org/10.1038/nm 1726) that modulation of replication fidelity is a promising approach for engineering attenuated virus vaccines. The subsequently mutagen-isolated RdRp variants also expressed the high-fidelity polymerase, but not all of them were attenuated. Few studies have shown the exact correlation between fidelity and virulence. The present study investigates the effect of restricted quasispecies diversity on viral virulence via several attenuated FMDV high-fidelity RdRp variants. Our findings may aid in the rational design of a new type of vaccine strain.

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Section: Generation Of Fmdv Populations Resistant To Ribavirinmentioning

confidence: 71%

mentioning

confidence: 62%

Section: Discussionmentioning

confidence: 99%

Section: Generation Of Fmdv Populations Resistant To Ribavirinmentioning

confidence: 99%

Section: Generation Of Fmdv Populations Resistant To Ribavirinmentioning

confidence: 99%

See 3 more Smart Citations

Ribavirin-Resistant Variants of Foot-and-Mouth Disease Virus: the Effect of Restricted Quasispecies Diversity on Viral Virulence

Zeng

Wang

Xie

et al. 2014

J Virol

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Fidelity Variants and RNA Quasispecies

Bordería

Rozen-Gagnon

Vignuzzi

2015

Current Topics in Microbiology and Immunology

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By now, it is well established that the error rate of the RNA-dependent RNA polymerase (RdRp) that replicates RNA virus genomes is a primary driver of the mutation frequencies observed in RNA virus populations-the basis for the RNA quasispecies. Over the last 10 years, a considerable amount of work has uncovered the molecular determinants of replication fidelity in this enzyme. The isolation of high- and low-fidelity variants for several RNA viruses, in an expanding number of viral families, provides evidence that nature has optimized the fidelity to facilitate genetic diversity and adaptation, while maintaining genetic integrity and infectivity. This chapter will provide an overview of what fidelity variants tell us about RNA virus biology and how they may be used in antiviral approaches.

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Foot-and-mouth disease virus low-fidelity polymerase mutants are attenuated

et al. 2014

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Previous studies have shown that RNA viruses can be attenuated by either increased or decreased viral polymerase replication fidelity. Although foot-and-mouth disease virus (FMDV) high-fidelity RNA-dependent RNA polymerase (RdRp) variants with an attenuated phenotype have been isolated using mutagens, no FMDV mutant with a low-fidelity polymerase has been documented to date. Here, we describe the generation of several FMDV RdRp mutants using site-directed mutagenesis via a reverse genetic system. Mutation frequency assays confirmed that five rescued FMDV RdRp mutant populations had lower replication fidelity than the wild-type virus population, which allowed us to assess the effects of the change in replication fidelity on the virus phenotype. These low-fidelity FMDV RdRp mutants showed increased sensitivity to ribavirin or 5-fluorouracil (5-FU) treatment without a loss of growth capacity in cell cultures. In addition, decreased fitness and attenuated virulence were observed for the RdRp mutants with lower fidelity. Importantly, based on a quantitative analysis for fidelity and virulence, we concluded that lower replication fidelity is associated with a more attenuated virus phenotype. These results further contribute to our understanding of the replication fidelity of polymerases of RNA viruses and its relationship to virulence attenuation.

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An increased replication fidelity mutant of foot-and-mouth disease virus retains fitness in vitro and virulence in vivo

Cited by 42 publications

References 36 publications

Ribavirin-Resistant Variants of Foot-and-Mouth Disease Virus: the Effect of Restricted Quasispecies Diversity on Viral Virulence

Ribavirin-Resistant Variants of Foot-and-Mouth Disease Virus: the Effect of Restricted Quasispecies Diversity on Viral Virulence

Fidelity Variants and RNA Quasispecies

Foot-and-mouth disease virus low-fidelity polymerase mutants are attenuated

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