Targeted mutagenesis of the herpesvirus fusogen central helix captures transition states.
Targeted mutagenesis of the herpesvirus fusogen central helix captures transition states.
复制标题
疱疹病毒融合剂中央螺旋的定向诱变捕获过渡态。
DOI：
10.1038/s41467-023-43011-w
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发表时间：
2023-12-02
影响因子：
16.6
通讯作者：
Oliver, Stefan L.
中科院分区：
文献类型：
--
作者：
Zhou, Momei;Vollmer, Benjamin;Machala, Emily;Chen, Muyuan;Gruenewald, Kay;Arvin, Ann M.;Chiu, Wah;Oliver, Stefan L.
疱疹病毒仍然是动物和人类健康的负担，包括医学上重要的水痘带状疱疹病毒(VZV)。由保守的核心糖蛋白FusoGen GB和异源二聚体Gh-gl介导的膜融合使疱疹病毒能够进入细胞。GB同源基因的胞外结构域有五个结构域，被认为是从融合前构象转换到融合后构象，但这一转换过程中结构域的功能相关性仍未得到很好的确定。在这里，我们描述了靶向残基526EHV528的VZV GB DIII中心螺旋的结构和功能研究。关键的是，H527P突变以冷冻-EM确定的预融合构象捕获GB，在无病毒检测中失去膜融合，以及重组VZV未能在细胞单层中传播。重要的是，通过三维分类和聚焦细化，确定了GB[H527P]的两个主要低温EM结构，表明它们代表了转变过程中的GB构象。这些研究表明，gB DIII是疱疹病毒gB融合功能的关键元件。包膜病毒感染细胞所需的病毒FusoGen构象变化的早期阶段尚不清楚。在这里，作者通过低温EM捕获了处于早期过渡状态的疱疹病毒FusoGen，即糖蛋白B。
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. The early stages of viral fusogen conformational change required for enveloped viruses to infect cells is unclear. Here, authors capture the herpesvirus fusogen, glycoprotein B, in early transitional states by cryo-EM.
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影响因子：
16.6
作者：
Krarup A;Truan D;Furmanova-Hollenstein P;Bogaert L;Bouchier P;Bisschop IJM;Widjojoatmodjo MN;Zahn R;Schuitemaker H;McLellan JS;Langedijk JPM
通讯作者：
Langedijk JPM
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
5.4
作者：
Jones, TR;Lee, SW;O'Connell, J
通讯作者：
O'Connell, J
影响因子：
7.2
作者：
Braakman, Ineke;Hebert, Daniel N.
通讯作者：
Hebert, Daniel N.
影响因子：
3.9
作者：
Oliver SL;Zhou M;Arvin AM
通讯作者：
Arvin AM