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

在病毒进入宿主细胞后，衣壳的分解和随后的病毒基因组的释放是SV 40感染的基本步骤，尽管其机制尚不清楚。当病毒颗粒缺乏次要衣壳蛋白Vp 2和Vp 3（统称为Vp 2/3）时，通常由细胞提示诱导的病毒分解没有得到适当的调节：缺乏Vp 2/3的颗粒在进入细胞时过早地释放病毒DNA（Nakanishi等人，J.Virol. 2007年）。为了研究Vp 1和Vp 2/3相互作用对病毒解体的调控作用，对Vp 1和Vp 2/3结合位点进行了突变，并分析了突变对病毒感染周期的影响。突变设计为（1）改变Vp 1和Vp 2/3、Vp 1 K116 R和Vp 3 Q174 D之间的离子相互作用界面，以及（2）在Vp 1和Vp 2/3、Vp 1 V230 C-Vp 3 E159 C和Vp 1 E257 C-Vp 3 Q174 C之间形成二硫键。此外，（3）一个突变体结合了三个不同的温度， ...更多信息 在分析中还包括了温度敏感突变（tsD），每一种都被报道表现出“未包被缺陷”，Vp 3 P103 S-M110 I-Q113 K。在转染突变病毒DNA后，衣壳蛋白表达和病毒DNA复制正常发生，并且所有突变体形成具有与野生型病毒体相似组成的颗粒。而在37 ℃或限制性温度下（Vp 3 P103 S-M110 I-Q113 K），除Vp 1 K116 R外，其余突变体均不能形成噬斑。突变体颗粒显然能够进入细胞，与importin α/β异二聚体相互作用，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。结果表明，病毒衣壳主要的构象变化涉及Vp 1-Vp 2/3相互作用的转移，这种变化发生在病毒进入细胞的后期，包括进入核后事件。少

项目成果

Molecular dissection of nuclear entry-competent SV40 during infection

• DOI: 10.1016/j.virusres.2006.10.001
• 发表时间: 2007-03-01
• 期刊: VIRUS RESEARCH
• 影响因子: 5
• 作者: Nakanishi, Akira;Li, Peggy P.;Kasamatsu, Harumi
• 通讯作者: Kasamatsu, Harumi

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.

Minor capsid proteins of SV40 are dispensable for nucleocapsid assembly and cell entry, but are required for nuclear entry of the viral genome

SV40 的次要衣壳蛋白对于核衣壳组装和细胞进入是可有可无的，但对于病毒基因组的核进入是必需的

• 发表时间: 2007
• 期刊: J.Virol. 81
• 作者: Nakanishi A;Itoh N;Li P P;Handa H;Liddington R C;Kasamatsu H
• 通讯作者: Kasamatsu H

The VP2/VP3 minor capsid protein of simian virus 40 promotes the in vitro assembly of the major capsid protein VP1 into particles

• DOI: 10.1074/jbc.m511261200
• 发表时间: 2006-04-14
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Kawano, M;Inoue, T;Handa, H
• 通讯作者: Handa, H