Amino acid substitutions contributing to α2,6‐sialic acid linkage binding specificity of human parainfluenza virus type 3 hemagglutinin–neuraminidase

Fukushima, Keijo; Takahashi, Toshifumi; Ueyama, Hiroo; Takaguchi, Masahiro; Itô, Seigo; Oishi, Kenta; Minami, Akira; Ishitsubo, Erika; Tokiwa, Hiroaki; Takimoto, Toru; Suzuki, Takashi

doi:10.1016/j.febslet.2015.03.036

Cited by 11 publications

(5 citation statements)

References 10 publications

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“…Further, our experiments with EV-D68 suggest that the entry of many, if not all, viruses that require sialic acid for entry would also be inhibited. Other viruses not specifically tested in this study but which we predict could be susceptible to B3GAT1 activity include important human pathogens such as the human coronaviruses OC43 and HKU-1 63 , parainfluenza virus types 1 and 3 64 , 65 and Mumps virus 66 . While much work remains before the clinical use of host-directed antiviral therapies, these data suggest that continued research in this area may eventually lead to novel and potentially truly broad-spectrum antivirals.…”

Section: Discussionmentioning

confidence: 99%

Cellular glycan modification by B3GAT1 broadly restricts influenza virus infection

et al. 2022

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Communicable respiratory viral infections pose both epidemic and pandemic threats and broad-spectrum antiviral strategies could improve preparedness for these events. To discover host antiviral restriction factors that may act as suitable targets for the development of host-directed antiviral therapies, we here conduct a whole-genome CRISPR activation screen with influenza B virus (IBV). A top hit from our screen, beta-1,3-glucuronyltransferase 1 (B3GAT1), effectively blocks IBV infection. Subsequent studies reveal that B3GAT1 activity prevents cell surface sialic acid expression. Due to this mechanism of action, B3GAT1 expression broadly restricts infection with viruses that require sialic acid for entry, including Victoria and Yamagata lineage IBVs, H1N1/H3N2 influenza A viruses (IAVs), and the unrelated enterovirus D68. To understand the potential utility of B3GAT1 induction as an antiviral strategy in vivo, we specifically express B3GAT1 in the murine respiratory epithelium and find that overexpression is not only well-tolerated, but also protects female mice from a lethal viral challenge with multiple influenza viruses, including a pandemic-like H1N1 IAV. Thus, B3GAT1 may represent a host-directed broad-spectrum antiviral target with utility against clinically relevant respiratory viruses.

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Section: Discussionmentioning

confidence: 99%

Cellular glycan modification by B3GAT1 broadly restricts influenza virus infection

et al. 2022

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“…Further validation of the method was carried out by comparing the results to those from several well-established assays of p7 activity. In case of the haemadsorption assay, which has been used by numerous groups, including ourselves [21,35,[48][49][50][51], we attempted to adapt the assay from its classical 6-well format [21,35,48,50] to a 24-well format [49,52] and make it suitable for quantitative analysis. However, cell damage due to p7 expression would lead to a loss of those cells that carry the recorded signal, and cell viability tests turned out to give more robust and reliable data.…”

Section: Discussionmentioning

confidence: 99%

Cell viability assay as a tool to study activity and inhibition of hepatitis C p7 channels

Breitinger

Farag

Ali

et al. 2021

Journal of General Virology

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The p7 viroporin of the hepatitis C virus (HCV) forms an intracellular proton-conducting transmembrane channel in virus-infected cells, shunting the pH of intracellular compartments and thus helping virus assembly and release. This activity is essential for virus infectivity, making viroporins an attractive target for drug development. The protein sequence and drug sensitivity of p7 vary between the seven major genotypes of the hepatitis C virus, but the essential channel activity is preserved. Here, we investigated the effect of several inhibitors on recombinant HCV p7 channels corresponding to genotypes 1a–b, 2a–b, 3a and 4a using patch-clamp electrophysiology and cell-based assays. We established a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based cell viability assay for recombinant p7 expressed in HEK293 cells to assess channel activity and its sensitivity to inhibitors. The results from the cell viability assay were consistent with control measurements using established assays of haemadsorption and intracellular pH, and agreed with data from patch-clamp electrophysiology. Hexamethylene amiloride (HMA) was the most potent inhibitor of p7 activity, but possessed cytotoxic activity at higher concentrations. Rimantadine was active against p7 of all genotypes, while amantadine activity was genotype-dependent. The alkyl-chain iminosugars NB-DNJ, NN-DNJ and NN-DGJ were tested and their activity was found to be genotype-specific. In the current study, we introduce cell viability assays as a rapid and cost-efficient technique to assess viroporin activity and identify channel inhibitors as potential novel antiviral drugs.

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“…Instead, the HPIV3 envelope glycoproteins, HN and F, form a fusion complex and work together to mediate viral attachment and entry into host cells. While the exact receptor is unknown, the HPIV3 transmembrane protein HN binds preferentially to a α2,3-linked sialic acid-containing receptor [ 4 , 5 ] on the host cell plasma membrane, and the F protein, once activated by the receptor-binding protein after receptor engagement [ 6 – 8 ], mediates the fusion of viral and host membranes, in order to deliver the viral genetic material into the host cell.…”

Section: Introductionmentioning

confidence: 99%

Human parainfluenza virus fusion complex glycoproteins imaged in action on authentic viral surfaces

et al. 2020

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Infection by human parainfluenza viruses (HPIVs) causes widespread lower respiratory diseases, including croup, bronchiolitis, and pneumonia, and there are no vaccines or effective treatments for these viruses. HPIV3 is a member of the Respirovirus species of the Paramyxoviridae family. These viruses are pleomorphic, enveloped viruses with genomes composed of single-stranded negative-sense RNA. During viral entry, the first step of infection, the viral fusion complex, comprised of the receptor-binding glycoprotein hemagglutininneuraminidase (HN) and the fusion glycoprotein (F), mediates fusion upon receptor binding. The HPIV3 transmembrane protein HN, like the receptor-binding proteins of other related viruses that enter host cells using membrane fusion, binds to a receptor molecule on the host cell plasma membrane, which triggers the F glycoprotein to undergo major conformational rearrangements, promoting viral entry. Subsequent fusion of the viral and host membranes allows delivery of the viral genetic material into the host cell. The intermediate states in viral entry are transient and thermodynamically unstable, making it impossible to understand these transitions using standard methods, yet understanding these transition states is important for expanding our knowledge of the viral entry process. In this study, we use cryoelectron tomography (cryo-ET) to dissect the stepwise process by which the receptor-binding protein triggers F-mediated fusion, when forming a complex with receptor-bearing membranes. Using an on-grid antibody capture method that facilitates examination of fresh, biologically active strains of virus directly from supernatant fluids and a series of biological tools that permit the capture of intermediate states in the fusion process, we visualize the series of events that occur when a pristine, authentic viral particle interacts with target receptors and proceeds from the viral entry steps of receptor engagement to membrane fusion.

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Amino acid substitutions contributing to α2,6‐sialic acid linkage binding specificity of human parainfluenza virus type 3 hemagglutinin–neuraminidase

Cited by 11 publications

References 10 publications

Cellular glycan modification by B3GAT1 broadly restricts influenza virus infection

Cellular glycan modification by B3GAT1 broadly restricts influenza virus infection

Cell viability assay as a tool to study activity and inhibition of hepatitis C p7 channels

Human parainfluenza virus fusion complex glycoproteins imaged in action on authentic viral surfaces

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