High resolution cryo EM analysis of HPV16 identifies minor structural protein L2 and describes capsid flexibility.
High resolution cryo EM analysis of HPV16 identifies minor structural protein L2 and describes capsid flexibility.
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DOI：
10.1038/s41598-021-83076-5
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发表时间：
2021-02-10
影响因子：
4.6
通讯作者：
Hafenstein SL
中科院分区：
文献类型：
--
作者：
Goetschius DJ;Hartmann SR;Subramanian S;Bator CM;Christensen ND;Hafenstein SL
人乳头瘤病毒（HPV）是一种重要的健康负担，也是病毒引起的癌症的主要原因。HPV是亲上皮的，并且其复制与终末角质形成细胞分化密切相关，使得用于研究的高滴度病毒制剂的生产和纯化成为问题，因此已经开发了替代的HPV生产方法用于病毒学和结构研究。在这项研究中，我们使用HPV 16准病毒，由HPV 16 L1/L2衣壳蛋白与包装的棉尾兔乳头瘤病毒基因组组成。我们已经实现了第一个高分辨率，3.1 μ m，结构的HPV 16通过使用局部子体积细化方法。高分辨率使我们能够明确地构建L1并识别L2蛋白链。L2密度被掺入到衣壳内部保守的L1残基附近。用我们自己的二十面体子体积提取和相关分类软件的进一步解释揭示了通过直径分析和用壳粒间分析的局部运动在整个颗粒水平上的灵活性。由连接臂支配的壳粒间膨胀或收缩在壳粒运动的幅度或方向上没有表现出偏差。我们建议，乳头瘤病毒衣壳是动态的和衣壳移动作为刚性机构连接灵活的链接。由此产生的病毒结构将为继续进行乳头瘤病毒的生物化学、遗传学和生物物理学研究提供框架。此外，我们的方法允许深入了解以前在乳头瘤病毒结构研究中限制的分辨率障碍。
Human papillomavirus (HPV) is a significant health burden and leading cause of virus-induced cancers. HPV is epitheliotropic and its replication is tightly associated with terminal keratinocyte differentiation making production and purification of high titer virus preparations for research problematic, therefore alternative HPV production methods have been developed for virological and structural studies. In this study we use HPV16 quasivirus, composed of HPV16 L1/L2 capsid proteins with a packaged cottontail rabbit papillomavirus genome. We have achieved the first high resolution, 3.1 Å, structure of HPV16 by using a local subvolume refinement approach. The high resolution enabled us to build L1 unambiguously and identify L2 protein strands. The L2 density is incorporated adjacent to conserved L1 residues on the interior of the capsid. Further interpretation with our own software for Icosahedral Subvolume Extraction and Correlated Classification revealed flexibility, on the whole-particle level through diameter analysis and local movement with inter-capsomer analysis. Inter-capsomer expansion or contraction, governed by the connecting arms, showed no bias in the magnitude or direction of capsomer movement. We propose that papillomavirus capsids are dynamic and capsomers move as rigid bodies connected by flexible linkers. The resulting virus structure will provide a framework for continuing biochemical, genetic and biophysical research for papillomaviruses. Furthermore, our approach has allowed insight into the resolution barrier that has previously been a limitation in papillomavirus structural studies.
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影响因子：
3.8
作者：
Brendle, Sarah A.;Culp, Timothy D.;Christensen, Neil D.
通讯作者：
Christensen, Neil D.
影响因子：
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Guan J;Bywaters SM;Brendle SA;Lee H;Ashley RE;Makhov AM;Conway JF;Christensen ND;Hafenstein S
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Hafenstein S
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通讯作者：
BROWN, JC
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影响因子：
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作者：
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通讯作者：
Trus, Benes L.