Recovery of infectious type Asia1 foot-and-mouth disease virus from suckling mice directly inoculated with an RNA polymerase I/II-driven unidirectional transcription plasmid

Lian, Kaiqi; Yang, Fan; Zhu, Zixiang; Cao, Weijun; Jin, Ye; Li, Dan; Zhang, Keshan; Guo, Jianpeng; Zheng, Haixue; Liu, Xiangtao

doi:10.1016/j.virusres.2015.06.008

Cited by 12 publications

(11 citation statements)

References 44 publications

(56 reference statements)

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“…For poliovirus, such clones are available by cloning of the poliovirus cDNA under transcriptional control of SV40-late promoter and SV40 termination signal [ 33 , 34 ]. The foot-and-mouth disease virus (FMDV) infectious plasmid DNA was transcriptionally controlled by both RNA polymerase I and II promoters which enabled production of high amounts of positive-stranded RNA [ 35 ]. An EV-A71 reverse genetic system under RNA polymerase I promoter was also reported previously [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

“…The presence of non-viral nucleotides at the 5’ and 3’ ends of enterovirus genomic RNA could reduce viral infectivity [ 16 , 25 , 45 , 46 ]. The HH ribozyme is essential for precise 5’ end generation [ 16 , 35 ]. The HH ribozyme belongs to a family of small endonucleolytic ribozymes that catalyze the cleavage of its own phosphodiester backbone by means of a trans-esterification reaction [ 47 , 48 ].…”

Section: Discussionmentioning

confidence: 99%

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Enterovirus A71 DNA-Launched Infectious Clone as a Robust Reverse Genetic Tool

et al. 2016

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Enterovirus A71 (EV-A71) causes major outbreaks of hand, foot and mouth disease, and is occasionally associated with neurological complications and death in children. Reverse genetics is widely used in the field of virology for functional study of viral genes. For EV-A71, such tools are limited to clones that are transcriptionally controlled by T7/SP6 bacteriophage promoter. This is often time-consuming and expensive. Here, we describe the development of infectious plasmid DNA-based EV-A71 clones, for which EV-A71 genome expression is under transcriptional control by the CMV-intermediate early promoter and SV40 transcriptional-termination signal. Transfection of this EV-A71 infectious DNA produces good virus yield similar to in vitro-transcribed EV-A71 infectious RNA, 6.4 and 5.8 log10PFU/ml, respectively. Infectious plasmid with enhanced green fluorescence protein and Nano luciferase reporter genes also produced good virus titers, with 4.3 and 5.0 log10 PFU/ml, respectively. Another infectious plasmid with both CMV and T7 promoters was also developed for easy manipulation of in vitro transcription or direct plasmid transfection. Transfection with either dual-promoter infectious plasmid DNA or infectious RNA derived from this dual-promoter clone produced infectious viral particles. Incorporation of hepatitis delta virus ribozyme, which yields precise 3’ ends of the DNA-launched EV-A71 genomic transcripts, increased infectious viral production. In contrast, the incorporation of hammerhead ribozyme in the DNA-launched EV-A71 resulted in lower virus yield, but improved the virus titers for T7 promoter-derived infectious RNA. This study describes rapid and robust reverse genetic tools for EV-A71.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Enterovirus A71 DNA-Launched Infectious Clone as a Robust Reverse Genetic Tool

et al. 2016

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“…The other three mutant viruses were generated based on prO. A DNA fragment, including the partial 5´UTR of FMDV O/BY/CHA/2010 with the 70-nt deletion as well as two restriction endonuclease enzyme sites, EcoRI and AfeI, was synthesized, and the fragment was placed into the prO as previously described (42) to generate pr-D70. pr-L10 was constructed with a similar strategy with KpnI and AfIII enzymes, and three nucleotides, TTG, were introduced into the L gene.…”

Section: Methodsmentioning

confidence: 99%

Genetic Determinants of Altered Virulence of Type O Foot-and-Mouth Disease Virus

Yang

Zhu

Cao

et al. 2020

J Virol

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Under different circumstances, the alteration of several viral genes may give an evolutionary advantage to the virus to maintain its prevalence in nature. In this study, a 70-nucleotide deletion in the small fragment (S fragment) of the viral 5′-untranslated region (5′-UTR) together with one amino acid insertion in the leader protein (Lpro) that naturally occurred in several serotype O foot-and-mouth disease virus (FMDV) strains in China was identified. The properties of two field serotype O FMDV strains, with or without the 70-nucleotide deletion in the S fragment and the amino acid insertion in Lpro, were compared in vitro and in vivo. Clinical manifestations of FMD were clearly observed in cattle and pigs infected by the virus without the mutations. However, the virus with the mentioned mutations caused FMD outcomes only in pigs, not in cattle. To determine the role of the 70-nucleotide deletion in the S fragment and the single amino acid insertion in Lpro in the pathogenicity and host range of FMDV, four recombinant viruses, with complete genomes and a 70-nucleotide deletion in the S fragment, a single amino acid insertion in Lpro, or both mutations, were constructed and rescued. It showed that deletion of 70 nucleotides in the S fragment or insertion of one amino acid (leucine) at position 10 of Lpro partly decreased the viral pathogenicity of Mya-98 lineage virus in cattle and pigs. However, the virus with dual mutations caused clinical disease only in pigs, not in cattle. This suggested that the S fragment and Lpro are significantly associated with the virulence and host specificity of FMDV. The naturally occurring dual mutation in the S fragment and Lpro is a novel determinant of viral pathogenicity and host range for serotype O FMDV. IMPORTANCE FMD is probably the most important livestock disease in the world due to the severe economic consequences caused. The alteration of several viral genes may give the virus selective advantage to maintain its prevalence in nature. Here, we identified that a 70-nucleotide deletion in the S fragment combined with a single leucine insertion in the leader protein (Lpro) is a novel determinant of restricted growth on bovine cells, which significantly contributes to the altered virulence of serotype O FMDV in cattle. A synergistic and additive effect of the 70-nucleotide deletion in the S fragment and the single leucine insertion in Lpro on the virulence and host specificity of the virus was determined. These results will benefit efforts to understand the vial pathogenicity mechanism and molecular characteristics of FMDV.

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“…Based on the reverse-genetics system established in our laboratory (37, 38), we constructed an infectious cDNA of O/CHA/99, termed pO-FMDV. The P1 coding sequence of O/BY/CHA/2010 was selected as the site for exchange with the corresponding region of pO-FMDV using the restriction endonucleases AfIII and Clal (New England Biolabs, Ipswich, Massachusetts, USA); the recombinant plasmid was named prVP3/FMDV.…”

Section: Methodsmentioning

confidence: 99%

The glycine locating at random coil of picornaviruses VP3 enhances viral pathogenicity by targeting p53 to promote apoptosis and autophagy

Mao¹,

Yang²,

Sun³

et al. 2019

Preprint

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Picornaviruses, comprising important and widespread pathogens of humans and animals, have evolved to control apoptosis and autophagy for their replication and spread. However, the underlying mechanism of the association between apoptosis/autophage and viral pathogenicity remains unclear. In the present study, VP3 of picornaviruses was demonstrated to induce apoptosis and autophagy. Foot-and-mouth disease virus (FMDV), which served as a research model here, can strongly induce both apoptosis and autophagy in the skin lesions. By directly interacting with p53, FMDV-VP3 facilitates its phosphorylation and translocation, resulting in Bcl-2 family-mediated apoptosis and LC3-dependent autophagy. The single residue Gly129 of FMDV-VP3 plays a crucial role in apoptosis and autophagy induction and the interaction with p53. Consistently, the comparison of rescued FMDV with mutated Gly129 and parental virus showed that the Gly129 is indispensable for viral replication and pathogenicity. More importantly, the Gly129 locates at a bend region of random coil structure, the mutation of Gly to Ala remarkably shrunk the volume of viral cavity. Coincidentally, the Gly is conserved in the similarly location of other picornaviruses, including poliovirus (PV), enterovirus 71 (EV71), coxsackievirus (CV) and seneca valley virus (SVA). This study demonstrates that picornaviruses induce apoptosis and autophagy to facilitate its pathogenicity and the Gly is functional site, providing novel insights into picornavirus biology.

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Recovery of infectious type Asia1 foot-and-mouth disease virus from suckling mice directly inoculated with an RNA polymerase I/II-driven unidirectional transcription plasmid

Cited by 12 publications

References 44 publications

Enterovirus A71 DNA-Launched Infectious Clone as a Robust Reverse Genetic Tool

Enterovirus A71 DNA-Launched Infectious Clone as a Robust Reverse Genetic Tool

Genetic Determinants of Altered Virulence of Type O Foot-and-Mouth Disease Virus

The glycine locating at random coil of picornaviruses VP3 enhances viral pathogenicity by targeting p53 to promote apoptosis and autophagy

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