The cap-binding site of influenza virus protein PB2 as a drug target

Severin, Chelsea; Moura, Tales Rocha de; Liu, Y.; Li, K.; Zheng, Xiaofeng; Luo, Ming

doi:10.1107/s2059798316000085

Cited by 25 publications

(58 citation statements)

References 15 publications

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“…Figure 1). Glu361 has one stable conformation and Lys376 has two, with only one suitable for hydrogen bonding to the guanine moiety of a cap analogue (Figure 1a) [10]. The sidechain of His357 is oriented out of plane but could rotate in plane to sandwich the guanine moiety together with the sidechain of Phe404 to achieve optimal π-force interactions.…”

Section: Data Processing and Refinementmentioning

confidence: 99%

“…The crystal structure of unliganded wild-type PB2cap from A/California/07/2009 (H1N1) (PDB code: 5EG8) [10] at 1.54 Å revealed the key residues available for interaction with the ligand ( Figure 1). Glu361 has one stable conformation and Lys376 has two, with only one suitable for hydrogen bonding to the guanine moiety of a cap analogue (Figure 1a) [10].…”

Section: Structure Of Pb2cap From A/california/07/2009 (H1n1) Comparementioning

confidence: 99%

“…The flexible sidechain of His432 and the dual-conforming sidechain of Asn429 could both participate in charge interactions with the α-phosphate in the cap. A water molecule is bound to the carbonyl of Phe404 that could easily be displaced upon ligand binding [10].…”

Section: Data Processing and Refinementmentioning

confidence: 99%

“…Table 1. Data collection, processing, structure solution, and refinement (values for the outer resolution shell are given in parentheses) [10,13]. …”

Section: Expression and Purificationmentioning

confidence: 99%

“…For the structure determination of the wild-type (PDB code: 5EG8), m 7 GTP-mutant (PDB code: 5EG7), and mutant CA09-PB2cap (PDB code: 5WOP) (Table 1), the A/California/07/2009 H1N1 DNA coding sequence was first inserted between NdeI and Xho1 restrictions sites into the pET28a expression vector to generate a His 6 -tag chimeric protein [10]. A QuickChange kit from Agilent Technologies, Santa Clara, CA, USA, was used to generate the expression vector for the mutant PB2cap.…”

Section: Expression and Purificationmentioning

confidence: 99%

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Characterization of the PB2 Cap Binding Domain Accelerates Inhibitor Design

et al. 2018

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X-ray crystallographic structural determinations of the PB2 cap binding domain (PB2cap) have improved the conformational characterization of the RNA-dependent RNA polymerase machinery (PA, PB2, and PB1) of the influenza virus. Geometrically, the catalytic PB1 subunit resembles the palm of a human hand. PA lies near the thumb region, and PB2 lies near the finger region. PB2 binds the cap moiety in the pre-mRNA of the host cell, while the endonuclease of PA cleaves the pre-mRNA 10-13 nucleotides downstream. The truncated RNA piece performs as a primer for PB1 to synthesize the viral mRNA. Precisely targeting PB2cap with a small molecule inhibitor will halt viral proliferation via interference of the cap-snatching activity. Wild-type and mutant PB2cap from A/California/07/2009 H1N1 were expressed in Escherichia coli, purified by nickel affinity and size exclusion chromatography, crystallized, and subjected to X-ray diffraction experiments. The crystal of mutant PB2cap liganded with m 7 GTP was prepared by co-crystallization. Structures were solved by the molecular replacement method, refined, and deposited in the Protein Data Bank (PDB). Structural determination and comparative analyses of these structures revealed the functions of Glu361, Lys376, His357, Phe404, Phe323, Lys339, His432, Asn429, Gln406, and Met401 in PB2cap, and the dissociation of the influenza A PB2cap C-terminal subdomain (residues 446-479) upon ligand binding. Understanding the role of these residues will aid in the ultimate development of a small-molecule inhibitor that binds both Influenza A and B virus PB2cap.

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Section: Data Processing and Refinementmentioning

confidence: 99%

Section: Structure Of Pb2cap From A/california/07/2009 (H1n1) Comparementioning

confidence: 99%

Section: Data Processing and Refinementmentioning

confidence: 99%

“…Table 1. Data collection, processing, structure solution, and refinement (values for the outer resolution shell are given in parentheses) [10,13]. …”

Section: Expression and Purificationmentioning

confidence: 99%

Section: Expression and Purificationmentioning

confidence: 99%

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Characterization of the PB2 Cap Binding Domain Accelerates Inhibitor Design

et al. 2018

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The Influenza Virus Polymerase Complex: An Update on Its Structure, Functions, and Significance for Antiviral Drug Design

Stevaert

Naesens

2016

Medicinal Research Reviews

137

151

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Influenza viruses cause seasonal epidemics and pandemic outbreaks associated with significant morbidity and mortality, and a huge cost. Since resistance to the existing anti‐influenza drugs is rising, innovative inhibitors with a different mode of action are urgently needed. The influenza polymerase complex is widely recognized as a key drug target, given its critical role in virus replication and high degree of conservation among influenza A (of human or zoonotic origin) and B viruses. We here review the major progress that has been made in recent years in unravelling the structure and functions of this protein complex, enabling structure‐aided drug design toward the core regions of the PA endonuclease, PB1 polymerase, or cap‐binding PB2 subunit. Alternatively, inhibitors may target a protein–protein interaction site, a cellular factor involved in viral RNA synthesis, the viral RNA itself, or the nucleoprotein component of the viral ribonucleoprotein. The latest advances made for these diverse pharmacological targets have yielded agents in advanced (i.e., favipiravir and VX‐787) or early clinical testing, besides several experimental inhibitors in various stages of development, which are all covered here.

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Multiple modes of action of myricetin in influenza A virus infection

Sang

Huang

Tian

et al. 2021

Phytotherapy Research

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Influenza A virus remains a major threat to public health worldwide after its first pandemic. Scientists keep searching novel anti-influenza drugs, of which natural products present to be an important source. Myricetin, a natural flavonol compound, which exists in many edible plants, which has a wide range of biological activities, but its anti-influenza A virus activity is ambiguous. This study aims to evaluate the antiinfluenza activity of myricetin and elucidate its underlying mechanism. Our results demonstrated that myricetin could significantly inhibit influenza A virus replication, reduce viral polymerase activity via selective inhibition of viral PB2 subunit, and the production of inflammatory cytokines by inhibiting TLR3 signaling pathway. The binding affinity analysis and the result of molecular docking revealed that myricetin interacted with the PB2 cap-binding pocket of influenza A virus. The above results suggested myricetin could exhibit anti-influenza virus activity with low cytotoxicity as well, and myricetin had low toxicity in BALB/c mice in vivo. Results from this study highlighted myricetin could be considered as a promising anti-influenza virus agent with dual inhibition profile. Furthermore, the compound with similar structure would provide a new option for the development of novel inhibitors against influenza A virus.

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The cap-binding site of influenza virus protein PB2 as a drug target

Cited by 25 publications

References 15 publications

Characterization of the PB2 Cap Binding Domain Accelerates Inhibitor Design

Characterization of the PB2 Cap Binding Domain Accelerates Inhibitor Design

The Influenza Virus Polymerase Complex: An Update on Its Structure, Functions, and Significance for Antiviral Drug Design

Multiple modes of action of myricetin in influenza A virus infection

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