A Conserved Domain in the Leader Proteinase of Foot-and-Mouth Disease Virus Is Required for Proper Subcellular Localization and Function

Santos, Teresa de los; Segundo, Fayna Diaz-San; Zhu, James; Koster, Marla; Dias, Cristina; Grubman, Marvin J.

doi:10.1128/jvi.02112-08

Cited by 64 publications

(95 citation statements)

References 48 publications

(56 reference statements)

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“…We have previously reported that FMDV lacking L pro is attenuated in swine and cattle; however, this virus is unable to completely protect animals against challenge with virulent FMDV (9,12,48). Recently, we constructed an FMDV mutant containing amino acid substitutions in a conserved domain of the L pro coding region, A12-SAP, which is attenuated in vitro (21). Here we show that A12-SAP-mutant FMDV is also attenuated in vivo.…”

Section: Discussionmentioning

confidence: 96%

“…In the case of FMDV, NF-B activation and translocation to the nucleus occur at a relatively early stage of infection, but at later times the p65/RelA subunit of NF-B disappears from infected cells (20). Previously, we demonstrated that FMDV L pro is necessary and sufficient for degradation of p65/RelA and that mutations in the L pro SAP domain abolished this function (20,21). Little is known about the influence of natural FMDV infection on the profile of proinflammatory cytokines in vivo.…”

Section: Discussionmentioning

confidence: 99%

“…Inhibition of IFN-␤ transcription is associated with L pro translocation to the nucleus of the infected cell and subsequent degradation of p65/RelA, a subunit of transcription factor nuclear factor kappa B (NF-B) (20). We have recently identified a conserved protein domain within the L pro coding region known as the SAF-A/B, acinus, and PIAS (SAP) domain (21). SAP domains which are present in some DNA binding proteins usually involved in transcriptional control mediate protein-protein interactions between activators and repressors (1,41).…”

mentioning

confidence: 99%

“…SAP domains which are present in some DNA binding proteins usually involved in transcriptional control mediate protein-protein interactions between activators and repressors (1,41). Mutations of two conserved amino acid residues in the SAP domain of FMDV L pro altered the protein subcellular localization during the course of infection, making the virus (SAP-mutant virus [A12-SAP]) unable to induce degradation of NF-B and thus resulting in upregulation of expression of several cytokines, chemokines, and interferon-stimulated genes (ISGs) (21).…”

mentioning

confidence: 99%

“…FMDV A12-WT was generated from the full-length serotype A12 infectious clone pRMC35 (66). A12-SAP-mutant virus, a derivative of A12-WT containing mutations I55A and L58A in the L pro region, was constructed by site-directed mutagenesis (21). All viruses were propagated in BHK-21 cells, concentrated by polyethylene glycol precipitation, titrated on the same cells, and stored at Ϫ70°C.…”

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confidence: 99%

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Inoculation of Swine with Foot-and-Mouth Disease SAP-Mutant Virus Induces Early Protection against Disease

Segundo

Weiss

Pérez-Martín

et al. 2012

J Virol

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Foot-and-mouth disease virus (FMDV) leader proteinase (L pro ) cleaves itself from the viral polyprotein and cleaves the translation initiation factor eIF4G. As a result, host cell translation is inhibited, affecting the host innate immune response. We have demonstrated that L pro is also associated with degradation of nuclear factor B (NF-B), a process that requires L pro nuclear localization. Additionally, we reported that disruption of a conserved protein domain within the L pro coding sequence, SAP mutation, prevented L pro nuclear retention and degradation of NF-B, resulting in in vitro attenuation. Here we report that inoculation of swine with this SAP-mutant virus does not cause clinical signs of disease, viremia, or virus shedding even when inoculated at doses 100-fold higher than those required to cause disease with wild-type (WT) virus. Remarkably, SAP-mutant virus-inoculated animals developed a strong neutralizing antibody response and were completely protected against challenge with WT FMDV as early as 2 days postinoculation and for at least 21 days postinoculation. Early protection correlated with a distinct pattern in the serum levels of proinflammatory cytokines in comparison to the levels detected in animals inoculated with WT FMDV that developed disease. In addition, animals inoculated with the FMDV SAP mutant displayed a memory T cell response that resembled infection with WT virus. Our results suggest that L pro plays a pivotal role in modulating several pathways of the immune response. Furthermore, manipulation of the L pro coding region may serve as a viable strategy to derive live attenuated strains with potential for development as effective vaccines against foot-and-mouth disease.

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Inoculation of Swine with Foot-and-Mouth Disease SAP-Mutant Virus Induces Early Protection against Disease

Segundo

Weiss

Pérez-Martín

et al. 2012

J Virol

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RNA‐virus proteases counteracting host innate immunity

Lei

Hilgenfeld

2017

FEBS Letters

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Virus invasion triggers host immune responses, in particular, innate immune responses. Pathogen‐associated molecular patterns of viruses (such as dsRNA, ssRNA, or viral proteins) released during virus replication are detected by the corresponding pattern‐recognition receptors of the host, and innate immune responses are induced. Through production of type‐I and type‐III interferons as well as various other cytokines, the host innate immune system forms the frontline to protect host cells and inhibit virus infection. Not surprisingly, viruses have evolved diverse strategies to counter this antiviral system. In this review, we discuss the multiple strategies used by proteases of positive‐sense single‐stranded RNA viruses of the families Picornaviridae, Coronaviridae, and Flaviviridae, when counteracting host innate immune responses.

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Modification of the second translation initiation site restricts the replication of foot-and-mouth disease virus in PK-15 cells

Yuan

et al. 2020

Appl Microbiol Biotechnol

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The translation initiation of foot-and-mouth disease virus (FMDV) occurs at two alternative initiation sites (Lab AUG and Lb AUG). Usually, the Lb AUG is more favorably used to initiate protein synthesis than the Lab AUG. To explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, this initiation codon was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA). Fortunately, the modifications did not prevent viral viability but influenced replication characteristics of some FMDV mutants in a cell-specific manner, as was shown by the similar replication in BHK-21 cells and delayed growth kinetics in PK-15 cells. This attenuated phenotype of FMDV mutants in PK-15 cells was found to be correlated with reduced abilities to cleave eIF4GI and suppress interference (IFN) expression. As leader (L) protein was reported to be responsible for eIF4GI cleavage and inhibition of IFN expression, the in vivo L protein synthesis was examined during the infection of FMDV mutants. Our results showed that not only the total yield of L proteins was severely influenced but also the individual yield of L protein was seen to be affected, which implied that both the relative usage of the two initiation sites and overall translation efficiency were changed by Lb AUG modifications. In addition, the in vitro translation activity was also negatively regulated by Lb AUG mutations. Collectively, these findings suggested that the restricted replications of Lb AUG-modified FMDVs were related to the delayed eIF4GI cleavage and decreased ability to block IFN expression but were mainly determined by the inefficient translation initiation. FMDVs precisely with modifications of Lb AUG initiation codon may represent safer seed viruses for vaccine production. Key points • The polyprotein translation of FMDV initiates at two alternative initiation sites (Lab AUG and Lb AUG). In order to explore the effect of Lb AUG on FMDV replication and obtain FMDV with restricted replication, the Lb initiation AUG was mutated to a variety of non-AUG codons (UGG, AUC, CUG, and AAA), and four FMDV mutants with Lb AUG modification were generated. • We found that partial FMDV mutants grew almost as well as WT virus in BHK-21 cells, a typical cell line used for FMD vaccine production, but displayed impaired replication in IFN-competent PK-15 cells. • The attenuation of mutant FMDVs in PK-15 cells was found to be correlated with delayed eIF4GI cleavage and decreased ability to block IFN expression. • We proved that the attenuated phenotype of Lb AUG-modified FMDVs was mainly determined by the inefficient translation initiation, as demonstrated by the decrease of total yield of L proteins and individual production of L protein. • We successfully generated genetically engineered FMDV with attenuated phenotype. The approach of precise engineering of FMDV with the modification of initiation codon provides a safe platform to produce inactivated antigen vaccines.

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A Conserved Domain in the Leader Proteinase of Foot-and-Mouth Disease Virus Is Required for Proper Subcellular Localization and Function

Cited by 64 publications

References 48 publications

Inoculation of Swine with Foot-and-Mouth Disease SAP-Mutant Virus Induces Early Protection against Disease

Inoculation of Swine with Foot-and-Mouth Disease SAP-Mutant Virus Induces Early Protection against Disease

RNA‐virus proteases counteracting host innate immunity

Modification of the second translation initiation site restricts the replication of foot-and-mouth disease virus in PK-15 cells

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