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

病毒进入囊膜的宿主细胞分解后，病毒基因组的随后释放是SV40感染的基本步骤，尽管其定义不明的机制是基础的。当病毒颗粒缺少较小的衣壳蛋白VP2和VP3（统称为VP2/3）时，病毒拆卸通常由细胞提示诱导，无法适当调节：VP2/3-无颗粒过早释放了病毒DNA（Nakanishi等人J.Virol。2007）。为了检查VP1和VP2/3相互作用对病毒拆卸调节的贡献，将突变引入了涉及VP1和VP2/3结合的残差，并分析了它们对病毒感染周期的影响。该突变旨在（1）改变VP1和VP2/3，VP1 K116R和VP3 Q174D之间的离子相互作用接口，并在VP1和VP2/3之间，VP1 V230C-VP3 E159C和VP1 E157C和VP1 E257C-VP3 Q174C之间形成二硫键。此外，（3）与三个不同的温度相结合的突变体……更敏感的敏感突变（TSD），据报道杀死“脱落缺陷”，VP3 P13S-M110I-Q113K，也包括在分析中。在转化突变的病毒DNA后，正常发生衣壳蛋白表达和病毒DNA复制，所有突变体形成与野生型病毒体相似组成的颗粒。但是，除VP1 K116R以外的突变体在37C或在限制性温度下形成斑块的能力严重有缺陷（VP3 P103S-M110I-Q113K）。突变颗粒显然能够进入细胞，与intimin alpha/beta异二聚体相互作用，尽管在启动病毒早期基因表达时有缺陷或延迟，表明突变体与病毒颗粒的核进口机械相互作用后，突变体的缺陷存在。结果表明，涉及VP1-VP2/3相互作用的病毒包囊的主要会议变化发生在细胞进入过程的后期，包括在感染期间，包括核后进入事件。较少的

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.