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TitleCryo-EM structure of the vaccinia virus entry fusion complex reveals a multicomponent fusion machinery.
Journal, issue, pagesSci Adv, Vol. 12, Issue 3, Page eaec0254, Year 2026
Publish dateJan 16, 2026
AuthorsChang Sheng-Huei Lin / Ching-An Li / Chun-Hsiung Wang / Chi-Fei Kao / Hsiao-Jung Chiu / Min-Chi Yeh / Hua-De Gao / Meng-Chiao Ho / Hsien-Ming Lee / Wen Chang
PubMed AbstractMembrane fusion is essential for viral entry. Unlike class I-III fusion proteins, vaccinia virus (VACV) uses a multicomponent entry fusion complex (EFC). Using cryo-electron microscopy, we determined ...Membrane fusion is essential for viral entry. Unlike class I-III fusion proteins, vaccinia virus (VACV) uses a multicomponent entry fusion complex (EFC). Using cryo-electron microscopy, we determined the full-length structure of the VACV EFC at near-atomic resolution, revealing a 15-protein asymmetric assembly organized into three layers. The central A16/G9/J5 heterotrimer forms the fusion core, stabilized by conserved PXXCW and Delta motifs, and anchors two A28/H2 adaptor dimers linked to peripheral G3/L5/A21/O3 scaffolds. Structural and evolutionary analyses identify a conserved N-terminal domain in A16 containing a myristoyl-binding pocket and a phenylalanine-rich region that stabilizes the trimer and may regulate lipid engagement. An additional component, F9, binds peripherally to J5, A21, and H2 through Delta-like motifs, reinforcing the prefusion architecture. Together, these results define the VACV EFC as a unique multiprotein fusion machinery and provide a structural framework for understanding the mechanism of poxvirus entry and membrane fusion.
External linksSci Adv / PubMed:41533782 / PubMed Central
MethodsEM (single particle)
Resolution3.05 Å
Structure data

EMDB-64647, PDB-9uzo:
Cryo-EM Structure of the Vaccinia Virus Entry/Fusion Complex (EFC) Lacking the F9 Subunit
Method: EM (single particle) / Resolution: 3.05 Å

EMDB-64648, PDB-9uzp:
Cryo-EM Structure of the Vaccinia Virus Entry/Fusion Complex (EFC) Including the F9 Subunit
Method: EM (single particle) / Resolution: 3.05 Å

Source
  • orthopoxvirus vaccinia
KeywordsVIRAL PROTEIN / Vaccinia Virus / Entry-Fusion Complex (EFC) / cryo-EM / Single-particle reconstruction

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