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TitleCryogenic electron microscopy and tomography reveal imperfect icosahedral symmetry in alphaviruses.
Journal, issue, pagesPNAS Nexus, Vol. 3, Issue 3, Page pgae102, Year 2024
Publish dateMar 7, 2024
AuthorsDavid Chmielewski / Guan-Chin Su / Jason T Kaelber / Grigore D Pintilie / Muyuan Chen / Jing Jin / Albert J Auguste / Wah Chiu /
PubMed AbstractAlphaviruses are spherical, enveloped RNA viruses with two-layered icosahedral architecture. The structures of many alphaviruses have been studied using cryogenic electron microscopy (cryo-EM) ...Alphaviruses are spherical, enveloped RNA viruses with two-layered icosahedral architecture. The structures of many alphaviruses have been studied using cryogenic electron microscopy (cryo-EM) reconstructions, which impose icosahedral symmetry on the viral particles. Using cryogenic electron tomography (cryo-ET), we revealed a polarized symmetry defect in the icosahedral lattice of Chikungunya virus (CHIKV) in situ, similar to the late budding particles, suggesting the inherent imperfect symmetry originates from the final pinch-off of assembled virions. We further demonstrated this imperfect symmetry is also present in in vitro purified CHIKV and Mayaro virus, another arthritogenic alphavirus. We employed a subparticle-based single-particle analysis protocol to circumvent the icosahedral imperfection and boosted the resolution of the structure of the CHIKV to ∼3 Å resolution, which revealed detailed molecular interactions between glycoprotein E1-E2 heterodimers in the transmembrane region and multiple lipid-like pocket factors located in a highly conserved hydrophobic pocket. This complementary use of in situ cryo-ET and single-particle cryo-EM approaches provides a more precise structural description of near-icosahedral viruses and valuable insights to guide the development of structure-based antiviral therapies against alphaviruses.
External linksPNAS Nexus / PubMed:38525304 / PubMed Central
MethodsEM (single particle) / EM (subtomogram averaging)
Resolution3.09 - 9.5 Å
Structure data

28979

8fcg

EMDB-28979, PDB-8fcg:
Cryo-EM structure of Chikungunya virus asymmetric unit
Method: EM (single particle) / Resolution: 3.09 Å

EMDB-41096: Cryo-electron tomography of Chikungunya virus pentamer structure
Method: EM (subtomogram averaging) / Resolution: 7.2 Å

EMDB-41631: Cryo-EM structure of Chikungunya virus with asymmetric reconstruction
Method: EM (single particle) / Resolution: 6.6 Å

EMDB-41637: Asymmetric cryoEM reconstruction of Mayaro virus
Method: EM (single particle) / Resolution: 9.5 Å

Chemicals

ChemComp-PCW:
1,2-DIOLEOYL-SN-GLYCERO-3-PHOSPHOCHOLINE / DOPC, phospholipid*YM

Source
  • chikungunya virus
KeywordsVIRUS / CHIKV asymmetric unit

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