Cellular DNAJA3, a Novel VP1-Interacting Protein, Inhibits Foot-and-Mouth Disease Virus Replication by Inducing Lysosomal Degradation of VP1 and Attenuating Its Antagonistic Role in the Beta Interferon Signaling Pathway

ABSTRACTDnaJ heat shock protein family (Hsp40) member A3 (DNAJA3) plays an important role in viral infections. However, the role of DNAJA3 in replication of foot-and-mouth-disease virus (FMDV) remains unknown. In this study, DNAJA3, a novel binding partner of VP1, was identified using yeast two-hybrid screening. The DNAJA3-VP1 interaction was further confirmed by coimmunoprecipitation and colocalization in FMDV-infected cells. The J domain of DNAJA3 (amino acids 1 to 168) and the lysine at position 208 (K208) of VP1 were shown to be critical for the DNAJA3-VP1 interaction. Overexpression of DNAJA3 dramatically dampened FMDV replication, whereas loss of function of DNAJA3 elicited opposing effects against FMDV replication. Mechanistical study demonstrated that K208 of VP1 was critical for reducing virus titer caused by DNAJA3 using K208A mutant virus. DNAJA3 induced lysosomal degradation of VP1 by interacting with LC3 to enhance the activation of lysosomal pathway. Meanwhile, we discovered that VP1 suppressed the beta interferon (IFN-β) signaling pathway by inhibiting the phosphorylation, dimerization, and nuclear translocation of IRF3. This inhibitory effect was considerably boosted in DNAJA3-knockout cells. In contrast, overexpression of DNAJA3 markedly attenuated VP1-mediated suppression on the IFN-β signaling pathway. Poly(I⋅C)-induced phosphorylation of IRF3 was also decreased in DNAJA3-knockout cells compared to that in the DNAJA3-WT cells. In conclusion, our study described a novel role for DNAJA3 in the host’s antiviral response by inducing the lysosomal degradation of VP1 and attenuating the VP1-induced suppressive effect on the IFN-β signaling pathway.IMPORTANCEThis study pioneeringly determined the antiviral role of DNAJA3 in FMDV. DNAJA3 was found to interact with FMDV VP1 and trigger its degradation via the lysosomal pathway. In addition, this study is also the first to clarify the mechanism by which VP1 suppressed IFN-β signaling pathway by inhibiting the phosphorylation, dimerization, and nuclear translocation of IRF3. Moreover, DNAJA3 significantly abrogated VP1-induced inhibitive effect on the IFN-β signaling pathway. These data suggested that DNAJA3 plays an important antiviral role against FMDV by both degrading VP1 and restoring of IFN-β signaling pathway.

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Inhibition of L-Deleted Foot-and-Mouth Disease Virus Replication by Alpha/Beta Interferon Involves Double-Stranded RNA-Dependent Protein Kinase

Journal of Virology ◽

10.1128/jvi.75.12.5498-5503.2001 ◽

2001 ◽

Vol 75 (12) ◽

pp. 5498-5503 ◽

Cited By ~ 105

Author(s):

Jarasvech Chinsangaram ◽

Marla Koster ◽

Marvin J. Grubman

Keyword(s):

Protein Kinase ◽

Disease Virus ◽

Foot And Mouth Disease ◽

Dependent Protein Kinase ◽

Double Stranded Rna ◽

Beta Interferon ◽

Mouth Disease Virus ◽

Dependent Protein ◽

Alpha Beta ◽

Fmdv Replication

ABSTRACT We previously demonstrated that the ability of foot-and-mouth disease virus (FMDV) to form plaques in cell culture is associated with the suppression of alpha/beta interferon (IFN-α/β). In the present study, we used Escherichia coli-expressed porcine and bovine IFN-α or -β individually to demonstrate that each was equally effective in inhibiting FMDV replication. The block in FMDV replication appeared to be at the level of protein translation, suggesting a role for double-stranded RNA-dependent protein kinase (PKR). In support of these findings, treatment of porcine and bovine cells with 2-aminopurine, an inhibitor of PKR, increased the yield of virus 8.8- and 11.2-fold, respectively, compared to that in untreated infected cells. In addition, results of FMDV infection in mouse embryonic fibroblast cells derived from gene knockout mice lacking the gene for RNase L−/− or PKR−/− or both indicated an important role for PKR in the inhibition of FMDV replication.

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Cellular Vimentin Interacts with Foot-and-Mouth Disease Virus Nonstructural Protein 3A and Negatively Modulates Viral Replication

Journal of Virology ◽

10.1128/jvi.00273-20 ◽

2020 ◽

Vol 94 (16) ◽

Cited By ~ 2

Author(s):

Xueqing Ma ◽

Ying Ling ◽

Pinghua Li ◽

Pu Sun ◽

Yimei Cao ◽

...

Keyword(s):

Disease Virus ◽

Virus Replication ◽

Nonstructural Protein ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Amino Acid Residues ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Fmdv Replication

ABSTRACT Nonstructural protein 3A of foot-and-mouth disease virus (FMDV) is a partially conserved protein of 153 amino acids that is in most FMDVs examined to date, and it plays important roles in virus replication, virulence, and host range. To better understand the role of 3A during FMDV infection, we used coimmunoprecipitation followed by mass spectrometry to identify host proteins that interact with 3A in FMDV-infected cells. Here, we report that cellular vimentin is a host binding partner for 3A. The 3A-vimentin interaction was further confirmed by coimmunoprecipitation, glutathione S-transferase (GST) pull down, and immunofluorescence assays. Alanine-scanning mutagenesis indicated that amino acid residues 15 to 21 at the N-terminal region of the FMDV 3A are responsible for the interaction between 3A and vimentin. Using reverse genetics, we demonstrate that mutations in 3A that disrupt the interaction between 3A and vimentin are also critical for virus growth. Overexpression of vimentin significantly suppressed the replication of FMDV, whereas knockdown of vimentin significantly enhanced FMDV replication. However, chemical disruption of the vimentin network by acrylamide resulted in a significant decrease in viral yield, suggesting that an intact vimentin network is needed for FMDV replication. These results indicate that vimentin interacts with FMDV 3A and negatively regulates FMDV replication and that the vimentin-3A interaction is essential for FMDV replication. This study provides information that should be helpful for understanding the molecular mechanism of FMDV replication. IMPORTANCE Foot-and-mouth disease virus (FMDV) nonstructural protein 3A plays important roles in virus replication, host range, and virulence. To further understand the role of 3A during FMDV infection, identification of host cell factors that interact with FMDV 3A is needed. Here, we found that vimentin is a direct binding partner of FMDV 3A, and manipulation of vimentin has a negative effect on virus replication. We also demonstrated that amino acid residues 15 to 21 at the N-terminal region of the FMDV 3A are responsible for the interaction between 3A and vimentin and that the 3A-vimentin interaction is critical for viral replication since the full-length cDNA clone harboring mutations in 3A, which were disrupt 3A-vimentin reactivity, could not produce viable virus progeny. This study provides information that not only provides us a better understanding of the mechanism of FMDV replication but also helps in the development of novel antiviral strategies in the future.

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The Leader Proteinase of Foot-and-Mouth Disease Virus Inhibits the Induction of Beta Interferon mRNA and Blocks the Host Innate Immune Response

Journal of Virology ◽

10.1128/jvi.80.4.1906-1914.2006 ◽

2006 ◽

Vol 80 (4) ◽

pp. 1906-1914 ◽

Cited By ~ 146

Author(s):

Teresa de los Santos ◽

Sonia de Avila Botton ◽

Rudi Weiblen ◽

Marvin J. Grubman

Keyword(s):

Disease Virus ◽

Foot And Mouth Disease ◽

Mrna Translation ◽

Mouth Disease ◽

Wild Type Virus ◽

Β Protein ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Fmdv Replication

ABSTRACT We have previously shown that the leader proteinase (Lpro) of foot-and-mouth disease virus (FMDV) blocks cap-dependent mRNA translation and that a genetically engineered FMDV lacking the leader proteinase coding region (A12-LLV2) is attenuated in cell culture and susceptible animals. The attenuated phenotype apparently is a consequence of the inability of A12-LLV2 to block the expression of type I interferon (IFN-α/β) protein, resulting in IFN-induced inhibition of FMDV replication. Here we show that in addition to preventing IFN-α/β protein synthesis, Lpro reduces the level of immediate-early induction of IFN-β mRNA and IFN-stimulated gene products such as double-stranded RNA-dependent protein kinase R (PKR), 2′,5′-oligoadenylate synthetase, and Mx1 mRNAs in swine cells. Down-regulation of cellular PKR by RNA interference did not affect wild-type virus yield but resulted in a higher yield of A12-LLV2, indicating a direct role of PKR in controlling FMDV replication in the natural host. The observation that Lpro controls the transcription of genes involved in innate immunity reveals a novel role of this protein in antagonizing the cellular response to viral infection.

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3Cpro of Foot-and-Mouth Disease Virus Antagonizes the Interferon Signaling Pathway by Blocking STAT1/STAT2 Nuclear Translocation

Journal of Virology ◽

10.1128/jvi.03668-13 ◽

2014 ◽

Vol 88 (9) ◽

pp. 4908-4920 ◽

Cited By ~ 49

Author(s):

Y. Du ◽

J. Bi ◽

J. Liu ◽

X. Liu ◽

X. Wu ◽

...

Keyword(s):

Signaling Pathway ◽

Disease Virus ◽

Nuclear Translocation ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Interferon Signaling ◽

Mouth Disease Virus ◽

Foot And Mouth

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Identifying the role of complement receptor 2 (CR2) on follicular dendritic cells (FDCs) in the persistence of foot and mouth disease virus (FMDV)

Access Microbiology ◽

10.1099/acmi.ac2019.po0074 ◽

2019 ◽

Vol 1 (1A) ◽

Author(s):

Lucy Gordon ◽

Neil Mabbott ◽

Bryan Charleston ◽

Eva Perez

Keyword(s):

Dendritic Cells ◽

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Complement Receptor ◽

Follicular Dendritic Cells ◽

Complement Receptor 2 ◽

Mouth Disease Virus ◽

Foot And Mouth

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Ability of Foot-and-Mouth Disease Virus To Form Plaques in Cell Culture Is Associated with Suppression of Alpha/Beta Interferon

Journal of Virology ◽

10.1128/jvi.73.12.9891-9898.1999 ◽

1999 ◽

Vol 73 (12) ◽

pp. 9891-9898 ◽

Cited By ~ 120

Author(s):

Jarasvech Chinsangaram ◽

Maria E. Piccone ◽

Marvin J. Grubman

Keyword(s):

Antiviral Activity ◽

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Host Cells ◽

Wild Type Virus ◽

Beta Interferon ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Alpha Beta

ABSTRACT A genetic variant of foot-and-mouth disease virus lacking the leader proteinase coding region (A12-LLV2) is attenuated in both cattle and swine and, in contrast to wild-type virus (A12-IC), does not spread from the initial site of infection after aerosol exposure of bovines. We have identified secondary cells from susceptible animals, i.e., bovine, ovine, and porcine animals, in which infection with A12-LLV2, in contrast to A12-IC infection, does not produce plaques; this result indicates that this virus cannot spread from the site of initial infection to neighboring cells. Nevertheless, A12-LLV2 can infect these cells, but cytopathic effects and virus yields are significantly reduced compared to those seen with A12-IC infection. Reverse transcription-PCR analysis demonstrates that both A12-LLV2 and A12-IC induce the production of alpha/beta interferon (IFN-α/β) mRNA in host cells. However, only supernatants from A12-LLV2-infected cells have significant antiviral activity. The antiviral activity in supernatants from A12-LLV2-infected embryonic bovine kidney cells is IFN-α/β specific, as assayed with mouse embryonic fibroblast cells with or without IFN-α/β receptors. The results obtained with cell cultures demonstrate that the ability of A12-IC to form plaques is associated with the suppression of IFN-α/β expression and suggest a role for this host factor in the inability of A12-LLV2 to spread and cause disease in susceptible animals.

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Lithium chloride inhibits early stages of foot-and-mouth disease virus (FMDV) replication in vitro

Journal of Medical Virology ◽

10.1002/jmv.24821 ◽

2017 ◽

Vol 89 (11) ◽

pp. 2041-2046 ◽

Cited By ~ 15

Author(s):

Fu-Rong Zhao ◽

Yin-Li Xie ◽

Ze-Zhong Liu ◽

Jun-Jun Shao ◽

Shi-Fang Li ◽

...

Keyword(s):

Disease Virus ◽

Lithium Chloride ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Early Stages ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Fmdv Replication

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The DDX23 Negatively Regulates Translation and Replication of Foot-and-Mouth Disease Virus and Is Degraded by 3C Proteinase

Viruses ◽

10.3390/v12121348 ◽

2020 ◽

Vol 12 (12) ◽

pp. 1348

Author(s):

Sahibzada Waheed Abdullah ◽

Shichong Han ◽

Jin’en Wu ◽

Yun Zhang ◽

Manyuan Bai ◽

...

Keyword(s):

Disease Virus ◽

Small Interfering Rna ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Dependent Manner ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Rna Knockdown ◽

Interfering Rna ◽

Fmdv Replication

DEAD-box helicase 23 (DDX23) is a host nuclear helicase, which is a part of the spliceosomal complex and involved in pre-mRNA splicing. To investigate whether DDX23, an internal ribosomal entry sites transacting factor (ITAF) affects foot-and-mouth disease virus (FMDV) replication and translation through internal ribosome entry site (IRES)-dependent manner. For this, we utilized a pull-down assay, Western blotting, quantitative real-time PCR, confocal microscopy, overexpression and small interfering RNA knockdown, as well as the median tissue culture infective dose. Our findings showed that FMDV infection inhibited DDX23 expression and the overexpression of DDX23 reduced viral replication, however, CRISPR Cas9 knockout/small interfering RNA knockdown increased FMDV replication. FMDV IRES domain III and IV interacted with DDX23, whereas DDX23 interacted with FMDV 3C proteinase and significantly degraded. The enzymatic activity of FMDV 3C proteinase degraded DDX23, whereas FMDV degraded DDX23 via the lysosomal pathway. Additionally, IRES-driven translation was suppressed in DDX23-overexpressing cells, and was enhanced in DDX23 knocked down. Collectively, our results demonstrated that DDX23 negatively affects FMDV IRES-dependent translation, which could be a useful target for the design of antiviral drugs.

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Basal Level p53 Suppresses Antiviral Immunity against Foot-and-Mouth Disease Virus

Viruses ◽

10.3390/v11080727 ◽

2019 ◽

Vol 11 (8) ◽

pp. 727

Author(s):

Zhang ◽

Chen ◽

Liu ◽

Qi ◽

Gao ◽

...

Keyword(s):

Innate Immunity ◽

Disease Virus ◽

Knockout Mice ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Clear Cut ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Immune Related Genes ◽

Fmdv Replication

Tumor suppressor protein p53 (p53) is a master transcription factor that plays key roles in cell cycle arrest, apoptosis, senescence, and metabolism, as well as regulation of innate immunity during virus infection. In order to facilitate their replication and spreading, viruses have evolved to manipulate p53 function through different strategies, with some requiring active p53 while others demand reduction/inhibition of p53 activity. However, there are no clear-cut reports about the roles of p53 during the infection of foot-and-mouth disease virus (FMDV), the causative agent of a highly contagious foot-and-mouth disease (FMD) of cloven-hoofed animals. Here we showed that p53 level was dynamically regulated during FMDV infection, being degraded at the early infection stage but recovered to the basal level at the late stage. Cells depleted of p53 showed inhibited FMDV replication and enhanced expression of the immune-related genes, whereas overexpression of p53 didn’t affect the viral replication. Viral challenge assay with p53 knockout mice obtained similar results, with viral load decreased, histopathological changes alleviated, and lifespan extended in the p53 knockout mice. Together, these data demonstrate that basal level p53 is required for efficient FMDV replication by suppressing the innate immunity.

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