Viroporin Activity of the Foot-and-Mouth Disease Virus Non-Structural 2B Protein

Ao, Da; Guo, Huichen; Sun, Shiqi; Sun, Dehui; Fung, To Sing; Wei, Yanquan; Han, Shuang; Yao, Xueping; Cao, Suizhong; Liu, Ding Xiang; Liu, Xiangtao

doi:10.1371/journal.pone.0125828

Cited by 49 publications

(73 citation statements)

References 65 publications

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“…7C). Following the pattern previously described to occur in Vero and BHK-21 cells (7,14), the wild-type fusion protein GFP (green fluorescent protein)-2B colocalized with the ER marker mCh-Sec61 beta in 293T cells. Mutants 2B.1, 2B.3, 2B.11, and 2B.13 displayed significantly lower Pearson colocalization indexes, indicating that alteration of the transmembrane structures interfered with the biogenesis of the 2B protein in these instances.…”

Section: Resultssupporting

confidence: 57%

“…FMDV 2B protein possesses ion channel activity. The FMDV 2B protein has been shown to behave as a viroporin regarding its cytotoxic effects upon overexpression in bacteria and its capacity for altering trafficking along early secretory pathways in animal cells, where it also seems to induce intracellular calcium release and autophagy (7). However, membrane permeability measurements investigating its pore formation mechanisms at the molecular level had not yet been performed.…”

Section: Resultsmentioning

confidence: 99%

“…It has been reported that the predicted structure of FMDV 2B contains two hydrophobic regions and inserts itself into the membrane of the ER with its N and C termini oriented toward the cytosol. The same report indicated that the 2B protein increases membrane permeability and can increase the Ca 2ϩ content in host cells, thereby inducing autophagy (7).…”

mentioning

confidence: 91%

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Molecular Characterization of the Viroporin Function of Foot-and-Mouth Disease Virus Nonstructural Protein 2B

Gladue

Largo

Arada

et al. 2018

J Virol

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Nonstructural protein 2B of foot-and-mouth disease (FMD) virus (FMDV) is comprised of a small, hydrophobic, 154-amino-acid protein. Structure-function analyses demonstrated that FMDV 2B is an ion channel-forming protein. Infrared spectroscopy measurements using partially overlapping peptides that spanned regions between amino acids 28 and 147 demonstrated the adoption of helical conformations in two putative transmembrane regions between residues 60 and 78 and between residues 119 and 147 and a third transmembrane region between residues 79 and 106, adopting a mainly extended structure. Using synthetic peptides, ion channel activity measurements in planar lipid bilayers and imaging of single giant unilamellar vesicles (GUVs) revealed the existence of two sequences endowed with membrane-porating activity: one spanning FMDV 2B residues 55 to 82 and the other spanning the C-terminal region of 2B from residues 99 to 147. Mapping the latter sequence identified residues 119 to 147 as being responsible for the activity. Experiments to assess the degree of insertion of the synthetic peptides in bilayers and the inclination angle adopted by each peptide regarding the membrane plane normal confirm that residues 55 to 82 and 119 to 147 of 2B actively insert as transmembrane helices. Using reverse genetics, a panel of 13 FMD recombinant mutant viruses was designed, which harbored nonconservative as well as alanine substitutions in critical amino acid residues in the area between amino acid residues 28 and 147. Alterations to any of these structures interfered with pore channel activity and the capacity of the protein to permeabilize the endoplasmic reticulum (ER) to calcium and were lethal for virus replication. Thus, FMDV 2B emerges as the first member of the viroporin family containing two distinct pore domains. FMDV nonstructural protein 2B is able to insert itself into cellular membranes to form a pore. This pore allows the passage of ions and small molecules through the membrane. In this study, we were able to show that both current and small molecules are able to pass though the pore made by 2B. We also discovered for the first time a virus with a pore-forming protein that contains two independent functional pores. By making mutations in our infectious clone of FMDV, we determined that mutations in either pore resulted in nonviable virus. This suggests that both pore-forming functions are independently required during FMDV infection.

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Section: Resultssupporting

confidence: 57%

Section: Resultsmentioning

confidence: 99%

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Molecular Characterization of the Viroporin Function of Foot-and-Mouth Disease Virus Nonstructural Protein 2B

Gladue

Largo

Arada

et al. 2018

J Virol

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“…A recent study demonstrated that 2B is a membrane-associated protein that acts as a viroporin and that 2B can induce intense autophagy in BHK-21 cells, thereby enhancing the release of viruses (48). The multiple alignment of the 2B coding sequences showed that it is highly conserved among all seven serotypes of FMDV (48).…”

Section: Discussionmentioning

confidence: 99%

“…The multiple alignment of the 2B coding sequences showed that it is highly conserved among all seven serotypes of FMDV (48). This implies that 2B contributes to the increased virus propagation and is associated with the infection mechanism of FMDV.…”

Section: Discussionmentioning

confidence: 99%

Foot-and-Mouth Disease Virus Viroporin 2B Antagonizes RIG-I-Mediated Antiviral Effects by Inhibition of Its Protein Expression

Zhu

Wang

Yang

et al. 2016

J Virol

125

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The role of retinoic acid-inducible gene I (RIG-I) in foot-and-mouth disease virus (FMDV)-infected cells remains unknown. Here, we showed that RIG-I inhibits FMDV replication in host cells. FMDV infection increased the transcription of RIG-I, while it decreased RIG-I protein expression. A detailed analysis revealed that FMDV leader proteinase (Lpro), as well as 3C proteinase (3Cpro) and 2B protein, decreased RIG-I protein expression. Lpro and 3Cpro are viral proteinases that can cleave various host proteins and are responsible for several of the viral polyprotein cleavages. However, for the first time, we observed 2B-induced reduction of host protein. Further studies showed that 2B-mediated reduction of RIG-I is specific to FMDV, but not other picornaviruses, including encephalomyocarditis virus, enterovirus 71, and coxsackievirus A16. Moreover, we found the decreased protein level of RIG-I is independent of the cleavage of eukaryotic translation initiation factor 4 gamma, the induction of cellular apoptosis, or the association of proteasome, lysosome, and caspase pathways. A direct interaction was observed between RIG-I and 2B. The carboxyl-terminal amino acids 105 to 114 and amino acids 135 to 144 of 2B were essential for the reduction of RIG-I, while residues 105 to 114 were required for the interaction. These data suggest the antiviral role of RIG-I against FMDV and a novel antagonistic mechanism of FMDV that is mediated by 2B protein.IMPORTANCE This study demonstrated that RIG-I could suppress FMDV replication during virus infection. FMDV infection increased the transcriptional expression of RIG-I, while it decreased RIG-I protein expression. FMDV 2B protein interacted with RIG-I and induced reduction of RIG-I. 2B-induced reduction of RIG-I was independent of the induction of the cleavage of eukaryotic translation initiation factor 4 gamma or cellular apoptosis. In addition, proteasome, lysosome, and caspase pathways were not involved in this process. This study provides new insight into the immune evasion mediated by FMDV and identifies 2B as an antagonistic factor for FMDV to evade the antiviral response.

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Cell pyroptosis in picornavirus and its potential for treating viral infection

Zheng

Zhuang

et al. 2022

Journal of Medical Virology

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Cell pyroptosis has received increased attention due to the associations between innate immunity and disease, and it has become a major focal point recently due to in‐depth studies of cancer. With increased research on pyroptosis, scientists have discovered that it has an essential role in viral infections, especially in the occurrence and development of some picornavirus infections. Many picornaviruses, including Coxsackievirus, a71 enterovirus, human rhinovirus, encephalomyocarditis virus, and foot‐and‐mouth disease virus induce pyroptosis to varying degrees. This review summarized the mechanisms by which these viruses induce cell pyroptosis, which can be an effective defense against pathogen infection. However, excessive inflammasome activation or pyroptosis also can damage the host's health or aggravate disease progression. Careful approaches that acknowledge this dual effect will aid in the exploration of picornavirus infections and the mechanisms that produce the inflammatory response. This information will promote the development of drugs that can inhibit cell pyroptosis and provide new avenues for future clinical treatment.

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Viroporin Activity of the Foot-and-Mouth Disease Virus Non-Structural 2B Protein

Cited by 49 publications

References 65 publications

Molecular Characterization of the Viroporin Function of Foot-and-Mouth Disease Virus Nonstructural Protein 2B

Molecular Characterization of the Viroporin Function of Foot-and-Mouth Disease Virus Nonstructural Protein 2B

Foot-and-Mouth Disease Virus Viroporin 2B Antagonizes RIG-I-Mediated Antiviral Effects by Inhibition of Its Protein Expression

Cell pyroptosis in picornavirus and its potential for treating viral infection

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