Mutagenesis of Human Cytomegalovirus Glycoprotein L Disproportionately Disrupts gH/gL/gO over gH/gL/pUL128-131
Mutagenesis of Human Cytomegalovirus Glycoprotein L Disproportionately Disrupts gH/gL/gO over gH/gL/pUL128-131
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人巨细胞病毒糖蛋白 L 的诱变不成比例地破坏 gH/gL/gO 而非 gH/gL/pUL128-131
DOI：
10.1128/jvi.00612-21
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发表时间：
2021
影响因子：
5.4
通讯作者：
Stegmann C
中科院分区：
文献类型：
--
作者：
Schultz EP;Stegmann C
疱疹病毒的无细胞传播和细胞间传播涉及由融合蛋白糖蛋白B（gB）和调节因子gH/gL组成的核心融合装置。人巨细胞病毒（HCMV）gH/gL/gO和gH/gL/pUL 128 -131促进在不同细胞类型中的传播。gO和pUL 128 -131组分结合不同的受体，但复合物的gH/gL部分在功能上如何比较尚不清楚。我们先前通过瞬时表达表征了一组gL突变体，并表明许多gH/gL-gB依赖性细胞-细胞融合受损，但仍能够形成gH/gL/pUL 128 -131并诱导受体干扰。在此，将gL突变体工程化到HCMV BAC克隆TB 40/e-BAC 4（TB）、TR和Merlin（ME）中，其在利用两种复合物进入和扩散方面不同。几个gL突变不成比例地影响gH/gL/gO依赖性进入并在gH/gL/pUL 128 -131过程中扩散。一些突变体的影响可以解释为受损的gH/gL/gO组装，但其他突变体影响gH/gL/gO功能。含有L201突变体的可溶性gH/gL/gO未能阻断HCMV感染，尽管与PDGFRα的结合未受损，表明存在其他重要的gH/gL/gO受体。另一个突变体（L139）增强TR的gH/gL/gO依赖性无细胞扩散，表明“过度活跃”的gH/gL/gO。最近发表的晶体学和冷冻电子显微镜研究表明，gH/gL/gO和gH/gL/pUL 128 -131的gH/gL的结构保守。然而，我们的数据表明，在两个复合物的gH/gL的重要差异，并支持一个模型，其中gH/gL/gO可以提供一个激活信号gB.IMPORTANCEThe地方性β疱疹病毒HCMV在人群中循环作为一个复杂的混合物的遗传不同的变种，建立终身持续感染，并导致重大疾病的新生儿和免疫功能低下的成年人。本研究利用我们最近对三种遗传上不同的HCMV BAC克隆的表征来辨别包膜糖蛋白复合物gH/gL/gO和gH/gL/pUL 128 -13的功能，这是共享疱疹病毒核心融合装置组分gH/gL的有希望的疫苗靶点。共享的gL亚基中的突变在功能上影响gH/gL/gO，证明了两种复合物之间的机制差异，并且可以为中和抗体的更精细评价提供基础。
Cell-free and cell-to-cell spread of herpesviruses involves a core fusion apparatus comprised of the fusion protein glycoprotein B (gB) and the regulatory factor gH/gL. The human cytomegalovirus (HCMV) gH/gL/gO and gH/gL/pUL128-131 facilitate spread in different cell types. The gO and pUL128-131 components bind distinct receptors, but how the gH/gL portions of the complexes functionally compare is not understood. We previously characterized a panel of gL mutants by transient expression and showed that many were impaired for gH/gL-gB-dependent cell-cell fusion but were still able to form gH/gL/pUL128-131 and induce receptor interference. Here, the gL mutants were engineered into the HCMV BAC clones TB40/e-BAC4 (TB), TR, and Merlin (ME), which differ in their utilization of the two complexes for entry and spread. Several of the gL mutations disproportionately impacted gH/gL/gO-dependent entry and spread over gH/gL/pUL128-131 processes. The effects of some mutants could be explained by impaired gH/gL/gO assembly, but other mutants impacted gH/gL/gO function. Soluble gH/gL/gO containing the L201 mutant failed to block HCMV infection despite unimpaired binding to PDGFRα, indicating the existence of other important gH/gL/gO receptors. Another mutant (L139) enhanced the gH/gL/gO-dependent cell-free spread of TR, suggesting a “hyperactive” gH/gL/gO. Recently published crystallography and cryo-electron microscopy studies suggest structural conservation of the gH/gL underlying gH/gL/gO and gH/gL/pUL128-131. However, our data suggest important differences in the gH/gL of the two complexes and support a model in which gH/gL/gO can provide an activation signal for gB.IMPORTANCEThe endemic betaherpesvirus HCMV circulates in human populations as a complex mixture of genetically distinct variants, establishes lifelong persistent infections, and causes significant disease in neonates and immunocompromised adults. This study capitalizes on our recent characterizations of three genetically distinct HCMV BAC clones to discern the functions of the envelope glycoprotein complexes gH/gL/gO and gH/gL/pUL128-13, which are promising vaccine targets that share the herpesvirus core fusion apparatus component, gH/gL. Mutations in the shared gL subunit disproportionally affected gH/gL/gO, demonstrating mechanistic differences between the two complexes, and may provide a basis for more refined evaluations of neutralizing antibodies.
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影响因子：
5.4
作者：
Paul T. Wille;A. J. Knoche;J. Nelson;M. Jarvis;David C. Johnson
通讯作者：
Paul T. Wille;A. J. Knoche;J. Nelson;M. Jarvis;David C. Johnson
影响因子：
5.4
作者：
Ryckman, BJ;Jarvis, MA;Johnson, DC
通讯作者：
Johnson, DC
DOI：
10.1073/pnas.0509201102
发表时间：
2005-12-13
影响因子：
11.1
作者：
Wang, D;Shenk, T
通讯作者：
Shenk, T
不由 UL128-131 和 RL13 位点决定的 BAC 克隆 HCMV 毒株的无细胞或细胞相关传播的专门化
DOI：
10.1101/760611
发表时间：
2019
期刊：
bioRxiv
影响因子：
--
作者：
E. Schultz;J. Lanchy;L. Z. Day;Q. Yu;Christopher Peterson;Jessica Preece;Brent J. Ryckman
通讯作者：
Brent J. Ryckman
DOI：
10.1099/vir.0.018994-0
发表时间：
2010-06
期刊：
The Journal of general virology
影响因子：
--
作者：
Dargan DJ;Douglas E;Cunningham C;Jamieson F;Stanton RJ;Baluchova K;McSharry BP;Tomasec P;Emery VC;Percivalle E;Sarasini A;Gerna G;Wilkinson GW;Davison AJ
通讯作者：
Davison AJ