Study of uncoating program of nuclear replicating virus

核复制病毒脱衣程序的研究

• 批准号: 17590416
• 负责人: NAKANISHI Akira
• 金额: $ 1.86万
• 依托单位: National Center for Geriatrics and Gerontology, National Institute for Longevity Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590416/
• 关键词: Virus; Capsid; Uncoating; Nuclear Import; virus structure; 構造変化; 感染; 自己組織化

Upon viral entry into the host cell disassembly of the capsid and subsequent release of the viral genome is the essential steps for SV40 infection, though mechanism underlies of which is poorly defined. The viral disassembly, normally induced by cellular cue, was not properly regulated when the viral particles lack the minor capsid proteins, Vp2 and Vp3 (collectively Vp2/3) : the Vp2/3-less particles prematurely released the viral DNA upon cell entry (Nakanishi et al. J.Virol. 2007). In order to examine the contribution of Vp1 and Vp2/3 interaction on the regulation of viral disassembly, mutations were introduced to the residues involving Vp1 and Vp2/3 binding, and analyzed their impact on viral infectious cycle. The mutations are designed to (1) alter the ionic interaction interface between Vp1 and Vp2/3, Vp1 K116R and Vp3 Q174D, and to (2) form disulfide bond between Vp1 and Vp2/3, Vp1 V230C-Vp3 E159C and Vp1 E257C-Vp3 Q174C. In addition (3) a mutant combined with three distinct temp … More erature sensitive mutations (tsDs), each reported to exhibit 'uncoating defect', Vp3 P103S-M110I-Q113K, was also included in the analysis. Upon transfection of the mutant viral DNA, capsid protein expression and viral DNA replication takes place normally, and all the mutants form particles with similar composition with that of wild-type virion. However, the mutants, except Vp1 K116R, were severely defective in their ability to form plaques at 37C or at the restrictive temperature (Vp3 P103S-M110I-Q113K). The mutant particles are apparently be able to enter the cells, interact with importin alpha/beta heterodimer, though either defective or delayed in initiating viral early gene expression, indicating that the defect of the mutants lies after interaction with nuclear import machinery of the viral particles. The results imply that major conformational change of viral capsid involving shift of Vp1-Vp2/3 interaction occurs at the later phase of cell entry processes, including the post-nuclear entry events, during the infection. Less

病毒进入宿主细胞后，衣壳的分解和随后病毒基因组的释放是 SV40 感染的基本步骤，尽管其机制尚不明确。病毒分解通常由细胞信号诱导，但在病毒分解时没有得到适当的调节。病毒颗粒缺乏次要衣壳蛋白 Vp2 和 Vp3（统称为 Vp2/3）：缺少 Vp2/3 的颗粒在进入细胞时过早释放病毒 DNA（Nakanishi 等） J.Virol. 2007）为了检查 Vp1 和 Vp2/3 相互作用对病毒解体调节的贡献，对涉及 Vp1 和 Vp2/3 结合的残基进行了突变，并分析了它们对病毒感染的影响。这些突变旨在 (1) 改变 Vp1 和 Vp2/3、Vp1 K116R 和 Vp3 Q174D 之间的离子相互作用界面，以及 (2)在 Vp1 和 Vp2/3、Vp1 V230C-Vp3 E159C 和 Vp1 E257C-Vp3 Q174C 之间形成二硫键。此外 (3) 与三个不同的温度敏感突变 (tsD) 结合的突变体，每个突变体均报告表现出“脱壳缺陷”。 '，Vp3 P103S-M110I-Q113K，也包含在分析中。突变体病毒DNA的转染、衣壳蛋白表达和病毒DNA复制均正常进行，并且所有突变体均形成与野生型病毒粒子组成相似的颗粒，但除Vp1 K116R外，突变体的能力存在严重缺陷。在 37°C 或限制温度下形成斑块 (Vp3 P103S-M110I-Q113K) 突变颗粒显然能够进入细胞，与细胞相互作用。 importin α/β异二聚体，尽管在启动病毒早期基因表达方面存在缺陷或延迟，表明突变体的缺陷在于与病毒颗粒的核输入机制相互作用之后。结果暗示涉及病毒衣壳转变的主要构象的变化。 Vp1-Vp2/3 相互作用发生在感染过程中细胞进入过程的后期，包括进入核后事件。

项目成果

Pairs of Vp1 cysteine residues essential for simian virus 40 infection

• DOI: 10.1128/jvi.79.6.3859-3864.2005
• 发表时间: 2005-03-01
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Li, PP;Nakanishi, A;Kasamatsu, H
• 通讯作者: Kasamatsu, H

The VP2/VP3 minor capsid protein of SV4O promotes the in vitro assembly of the major capsid protein VP1 into particles.

SV4O 的 VP2/VP3 次要衣壳蛋白促进主要衣壳蛋白 VP1 体外组装成颗粒。

• 发表时间: 2006
• 期刊: Journal of Biological Chemistry 281
• 作者: Kawano MA;Inoue T;Tsukamoto H;Takaya T;Enomoto T;Takahashi R;Yokoyama N;Yamamoto N;Nakanishi A;Imai T;Wada T;Kataoka K;Handa H.
• 通讯作者: Handa H.