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Cited by 13Poxvirus research and outbreaksVirology and Viral DiseasesProtein Structure and Dynamics

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Structural and functional analysis of vaccinia viral fusion complex component protein A28 through NMR and molecular dynamic simulations

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Kao, Chi-Fei, Tsai, Min-Hsin, Carillo, Kathleen Joyce, Tzou, Der-Lii, Chang, Wen·2023·PLOS Pathogens
DOI 10.1371/journal.ppat.1011500PMC PMC10664964

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Abstract

Host cell entry of vaccinia virus (a poxvirus) proceeds through multiple steps that involve many viral proteins to mediate cell infection. Upon binding to cells, vaccinia virus membrane fuses with host membranes via a viral entry fusion protein complex comprising 11 proteins: A16, A21, A28, F9, G3, G9, H2, J5, L1, L5 and O3. Despite vaccinia virus having two infectious forms, mature and enveloped, that have different membrane layers, both forms require an identical viral entry fusion complex for membrane fusion. Components of the poxvirus entry fusion complex that have been structurally assessed to date share no known homology with all other type I, II and III viral fusion proteins, and the large number of fusion protein components renders it a unique system to investigate poxvirus-mediated membrane fusion. Here, we determined the NMR structure of a truncated version of vaccinia A28 protein. We also expressed a soluble H2 protein and showed that A28 interacts with H2 protein at a 1:1 ratio in vitro . Furthermore, we performed extensive in vitro alanine mutagenesis to identify A28 protein residues that are critical for H2 binding, entry fusion complex formation, and virus-mediated membrane fusion. Finally, we used molecular dynamic simulations to model full-length A28-H2 subcomplex in membranes. In summary, we characterized vaccinia virus A28 protein and determined residues important in its interaction with H2 protein and membrane components. We also provide a structural model of the A28-H2 protein interaction to illustrate how it forms a 1:1 subcomplex on a modeled membrane.

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Metadata

Authors
Kao, C.-F., Tsai, M.-H., Carillo, K. J., Tzou, D.-L., & Chang, W.
Affiliations
  • Institute of Molecular Biology, Academia Sinica (TW)
  • Institute of Chemistry, Academia Sinica (TW)
Corresponding author
  • Tzou, Der-Lii
  • Chang, Wen
ORCID
Venue
PLOS Pathogens
Publisher
Public Library of Science (PLoS)
Published
2023-11-10
Volume
19
Issue
11
Pages
e1011500
ISSN
  • 1553-7366 (ISSN-L)
  • 1553-7374
DOI
10.1371/journal.ppat.1011500
PMC
PMC10664964
OpenAlex
W4388562933
Cited by
13
References
1
Citations by year
2026: 4 · 2025: 5 · 2024: 3 · 2023: 1
FWCI
1.71
Type
journal article
Language
en
Update notices
new_version (2023-11-22)
Open access
Licence
Locations
  • PLoS Pathogens (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Academia Sinica (AS-IDR-112-02)
  • National Science and Technology Council (110-2320-B-001 -015 -MY3, NSTC 111-2113-M-001-014)
  • Academia Sinica’s High-Field NMR Center (HFNMRC) Core Facility (AS-CFII-111-214)
  • Medicinal Chemistry and Analytical Core Facility (AS-NBRPCF-111-201)
Topics
  • Life Sciences › Immunology and Microbiology › Virology › Poxvirus research and outbreaks
  • Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
vaccinia virus, membrane fusion, poxvirus, protein-protein interaction, alanine scanning, NMR structure, molecular dynamics simulation, membrane protein complexes
MeSH terms
Humans, Poxviridae, Vaccinia, Vaccinia virus, Viral Fusion Proteins, Virus Internalization, Molecular Dynamics Simulation

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