Targeted mutagenesis of the herpesvirus fusogen central helix captures transition states

Zhou, Momei; Vollmer, Benjamin; Machala, Emily; Chen, Muyuan; Grünewald, Kay; Arvin, Ann M.; Chiu, Wah; Oliver, Stefan L.

doi:10.1038/s41467-023-43011-w

Targeted mutagenesis of the herpesvirus fusogen central helix captures transition states

Journal Article · Sat Dec 02 00:00:00 EST 2023 · Nature Communications

DOI:https://doi.org/10.1038/s41467-023-43011-w· OSTI ID:2405203

^[1]; ^[2]; ^[2]; Chen, Muyuan ^[3]; ^[2]; Arvin, Ann M. ^[4]; ^[5]; ^[6]

Stanford Univ., CA (United States); SLAC
Centre for Structural Systems Biology (CSSB), Hamburg (Germany); Univ. of Hamburg (Germany); Leibniz Institute of Virology (LIV), Hamburg (Germany)
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Stanford Univ., CA (United States); Vir Biotechnology Inc., San Francisco, CA (United States)
Stanford Univ., CA (United States); SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Stanford Univ., CA (United States)

Herpesviruses remain a burden for animal and human health, including the medically important varicella-zoster virus (VZV). Membrane fusion mediated by conserved core glycoproteins, the fusogen gB and the heterodimer gH-gL, enables herpesvirus cell entry. The ectodomain of gB orthologs has five domains and is proposed to transition from a prefusion to postfusion conformation but the functional relevance of the domains for this transition remains poorly defined. Here we describe structure-function studies of the VZV gB DIII central helix targeting residues ⁵²⁶EHV⁵²⁸. Critically, a H527P mutation captures gB in a prefusion conformation as determined by cryo-EM, a loss of membrane fusion in a virus free assay, and failure of recombinant VZV to spread in cell monolayers. Importantly, two predominant cryo-EM structures of gB[H527P] are identified by 3D classification and focused refinement, suggesting they represented gB conformations in transition. These studies reveal gB DIII as a critical element for herpesvirus gB fusion function.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: BMBF; DFG; National Institutes of Health (NIH); USDOE Office of Science (SC)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 2405203

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 14; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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Targeted mutagenesis of the herpesvirus fusogen central helix captures transition states

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