Recombinant MYH9 protein C-terminal domain blocks porcine reproductive and respiratory syndrome virus internalization by direct interaction with viral glycoprotein 5

Li, Liangliang; Xue, Biyun; Sun, Weihao; Gu, Guoqian; Hou, Gaopeng; Zhang, Lu; Wu, Chunyan; Zhao, Qin; Zhang, Yan‐Jin; Zhang, Gaiping; Hiscox, Julian A.; Nan, Yuchen; Zhou, En‐Min

doi:10.1016/j.antiviral.2018.06.001

Cited by 35 publications

(45 citation statements)

References 58 publications

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“…Theoretically, mutagenesis of above amino acid residues in PRA would impair binding of PRA mutants to virus, which reduce the inhibition effect of PRRSV by corresponding PRA mutants. To verify this, we first tested the PRA mutants developed here for an ELISA-based virus binding assay described before [35]. The results showed that all PRA mutants had significantly lower virus-binding activity than wild-type PRA ( Figure 6A).…”

Section: Impaired Binding Of Mab2-5g2 To Pra Mutants Correlates Withmentioning

confidence: 89%

“…Viruses were maintained in-house and used to inoculate MARC-145 cells or PAMs at the indicated multiplicity Viruses 2020, 12, 40 3 of 19 of infection (MOI) determined by evaluation of the median tissue culture infectious dose (TCID 50 ). Titration of different PRRSV strains was conducted in MARC-145 cells as previously described [35].…”

Section: Cells Viruses and Chemicalsmentioning

confidence: 99%

“…The specific interaction was detected using HRP-conjugated goat anti-rabbit IgG or anti-mouse IgG antibodies and visualized using a TMB kit (TianGen). In addition, PRRSV particle capture assays were further Viruses 2020, 12, 40 4 of 19 conducted to evaluate the interaction between PRA mutants and the PRRSV virion as previously described [35].…”

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

“…Wild-type PRA and truncated PRA were each cloned into pCold-SUMO vector followed by expression and purification of their corresponding proteins as previously described [35]. Briefly, the pCold-SUMO-PRA plasmid was transformed into the Escherichia coli strain BL21 (DE3) and cultured in LB medium at 37 • C until induced with 0.5 mM isopropyl β-d-thiogalactoside (IPTG) at 15 • C. After IPTG induction, bacterial cells were collected and resuspended in cell lysis buffer (50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM AEBSF (4-benzenesulfonyl fluoride hydrochloride), 5% glycerol) for sonication.…”

Section: Expression Of Pra Pra Truncation and Pra Mutantsmentioning

confidence: 99%

“…MYH9 has been identified as a cellular receptor for herpes simplex virus-1 (HSV-1) [31], severe fever with thrombocytopenia syndrome virus (SFTSV) [32], Epstein-Barr virus (EBV), and PRRSV [21,33]. Regarding PRRSV, the PRA domain located within the C-terminal portion of MYH9 is responsible for binding to viral GP5, as demonstrated using a recombinant soluble form of PRA that blocked PRRSV infection in vitro [34,35].…”

Section: Introductionmentioning

confidence: 99%

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MYH9 Key Amino Acid Residues Identified by the Anti-Idiotypic Antibody to Porcine Reproductive and Respiratory Syndrome Virus Glycoprotein 5 Involve in the Virus Internalization by Porcine Alveolar Macrophages

Zhang

et al. 2019

Viruses

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MYH9 has been identified as an indispensable cellular protein for porcine reproductive and respiratory syndrome virus (PRRSV) entry into permissive cells using the monoclonal anti-idiotypic antibody (Mab2-5G2) recognizing an antibody that specifically interacts with PRRSV glycoprotein 5 (GP5). More recently, we found that Mab2-5G2 interacted with the MYH9 C-terminal domain, designated PRA, which is required for PRRSV internalization. In this study, we demonstrate that blocking of MYH9 with Mab2-5G2 significantly diminished PRRSV internalization by porcine alveolar macrophage (PAM) via interruption of direct interaction between GP5 and MYH9, and thus remarkably inhibited subsequent infection of PAMs by PRRSV-2 isolates. Moreover, the three-dimensional structure of the Mab2-5G2 Fab-PRA complex determined via homology modeling predicted potential docking sites required for PRRSV internalization. Further analysis of Mab2-5G2-binding sites within PRA highlighted that the amino acids E1670, K1673, E1679, and I1683 in PRA are the key Mab2-5G2-binding residues. Notably, recombinant PRA protein blocked the interaction between PRRSV GP5 and cellular MYH9 by preventing translocation of MYH9 from the cytoplasm to the cell membrane, an essential step for PRRSV virion internalization. Meanwhile, porcine cell line permissive for PRRSV bearing point mutation of E1670A in MYH9 demonstrated reduced susceptibility for PRRSV infection. In conclusion, this work increases understanding of both PRRSV pathogenesis and the mechanistic role played by MYH9 in PRRSV infection.

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Section: Impaired Binding Of Mab2-5g2 To Pra Mutants Correlates Withmentioning

confidence: 89%

Section: Cells Viruses and Chemicalsmentioning

confidence: 99%

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

Section: Expression Of Pra Pra Truncation and Pra Mutantsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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MYH9 Key Amino Acid Residues Identified by the Anti-Idiotypic Antibody to Porcine Reproductive and Respiratory Syndrome Virus Glycoprotein 5 Involve in the Virus Internalization by Porcine Alveolar Macrophages

Zhang

et al. 2019

Viruses

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Myosin motors on the pathway of viral infections

2022

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Molecular motors are microscopic machines that use energy from adenosine triphosphate (ATP) hydrolysis to generate movement. While kinesins and dynein are molecular motors associated with microtubule tracks, myosins bind to and move on actin filaments. Mammalian cells express several myosin motors. They power cellular processes such as endo‐ and exocytosis, intracellular trafficking, transcription, migration, and cytokinesis. As viruses navigate through cells, they may take advantage or be hindered by host components and machinery, including the cytoskeleton. This review delves into myosins' cell roles and compares them to their reported functions in viral infections. In most cases, the previously described myosin functions align with their reported role in viral infections, although not in all cases. This opens the possibility that knowledge obtained from studying myosins in viral infections might shed light on new physiological roles for myosins in cells. However, given the high number of myosins expressed and the variety of viruses investigated in the different studies, it is challenging to infer whether the interactions found are specific to a single virus or can be applied to other viruses with the same characteristics. We conclude that the participation of myosins in viral cycles is still a largely unexplored area, especially concerning unconventional myosins.

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Broad neutralization activity against both PRRSV-1 and PRRSV-2 and enhancement of cell mediated immunity against PRRSV by a novel IgM monoclonal antibody

Zhai

et al. 2020

Antiviral Research

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Recombinant MYH9 protein C-terminal domain blocks porcine reproductive and respiratory syndrome virus internalization by direct interaction with viral glycoprotein 5

Cited by 35 publications

References 58 publications

MYH9 Key Amino Acid Residues Identified by the Anti-Idiotypic Antibody to Porcine Reproductive and Respiratory Syndrome Virus Glycoprotein 5 Involve in the Virus Internalization by Porcine Alveolar Macrophages

MYH9 Key Amino Acid Residues Identified by the Anti-Idiotypic Antibody to Porcine Reproductive and Respiratory Syndrome Virus Glycoprotein 5 Involve in the Virus Internalization by Porcine Alveolar Macrophages

Myosin motors on the pathway of viral infections

Broad neutralization activity against both PRRSV-1 and PRRSV-2 and enhancement of cell mediated immunity against PRRSV by a novel IgM monoclonal antibody

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