Complete Structural Determination of Rice dwarf virus

水稻矮缩病毒的完整结构测定

• 批准号: 14380319
• 负责人: NAKAGAWA Atsushi
• 金额: $ 9.41万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14380319/
• 关键词: Biological macromolecular assemblies; Synchrotron Radiation; Virus; Structure assembly of virus; X-ray crystal structure analysis; Infection mechanism of virus; Self-replication mechanism of virus; 多波長異常分散法; タンパク質複合体

This project has been worked on the structural studies of Rice dwarf virus, which is a double-shelled virus with its molecular mass of about 7,500 kDa, to reveal its whole structure using x-ray crystallography, cryo-electron micrograph and molecular biological techniques.Most of the structural studies have been done with the P2-deletion RDV particle, which contained all viral components except the P2 protein, where P2 protein had been removed from the sample during treatment with CCl_4 in the purification step. The 3.5Å resolution electron density map of RDV crystal, determined by x-ray crystallography, revealed almost complete structures of the inner- and outer-shells (P3 and P8 proteins) and a part of P7 protein. The interactions between the subunit proteins suggest how the 900 protein components are built into a higher-ordered virus core structure, and we proposed the model for the hierarchy of structural assembly. This model was strongly supported by some experiments using molecular biological techniques.Internal structure of the virus particle was studied by combination of the single-particle analysis using cryo-electron micrograph and x-ray crystallography. Low-resolution cryo-EM map showed clear molecular envelop corresponding to the transcription complex (complex of P1,P5 and P7 proteins). The map also clearly showed at lease three layered-structure of dsRNA. Especially, dsRNAs in the first and second layer showed rod-shaped structure, which are packed into liquid crystal-like structure.In addition, we have succeeded to purify the component proteins from the virus particle and/or from overexpressed sample, these samples were subjected to crystallization to reveal the high-resolution structure of the component proteins.

该项目已在大米矮人病毒的结构研究中进行过研究，该病毒是一种双壳病毒，其分子质量约为7,500 kDa，使用X射线晶体学，冷冻电子显微照片和分子生物学技术揭示其整个结构，其中最多可与P2-delet prect rdv rdv rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd rd。在纯化步骤中用CCL_4处理期间，已从样品中删除。由X射线晶体学确定的RDV晶体的3.5Å分辨率电子密度图揭示了内壳和外壳（P3和P8蛋白）的几乎完整结构和P7蛋白的一部分。亚基蛋白之间的相互作用表明900蛋白成分是如何内置在高阶病毒核心结构中的，我们提出了结构组装层次结构的模型。通过某些实验使用分子生物学技术，通过使用Cryo-Electron显微照片和X射线晶体学的单粒子分析研究了该模型。低分辨率的冷冻EM图显示了与转录复合物（P1，P5和P7蛋白的复合物）相对应的透明分子包膜。该地图还清楚地显示了DSRNA的三个分层结构。尤其是，在第一层和第二层中显示了棒状结构中的DSRNA，这些结构被包装到液晶状结构中。此外，我们成功地从病毒颗粒和/或从过表达的样品中净化了组分蛋白，这些样品被结晶以揭示组分蛋白的高分辨率结构。

项目成果

イネ萎縮ウイルスの立体構造

水稻萎缩病毒三维结构

• 发表时间: 2004
• 期刊: BioMedical Quick Review Net 4028
• 作者: Aki Nagata;Aki Nagata;Aki Nagata;Tsutomu Nakamura;Noriko Higashi;Kazuko Fujiwara;Kazuko Okamura-Ikeda;Shu Jie Li;Junichiro Taka;Kazunori Namba;Shu Jie Li;Sachiko Toma;Naoyuki Miyazaki;Hiroyuki Akama;赤間浩之;大村敏博;Tsutomu Nakamura;Noriko Higashi;Kazuko Okamura-Ikeda;Junichiro Taka;Shu Jie Li;Naoyuki Miyazaki;Hiroyuki Akama;Junichiro Taka;Tsutomu Nakamura;Noriko Higashi;Kazuko Fujiwara;Kazuko Okamura-Ikeda;Shu Jie Li;Shu Jie Li;Hiroyuki Akama;N.Miyazaki;S.Toma;Hiroshi Sasaki;Hiroyuki Akama;S.J.Li;S.Bando;T.Uchida;H.Akama;J.Ma;J.Ma;Kyoji Hagiwara;内藤久志;中川敦史;宮崎直幸
• 通讯作者: 宮崎直幸

ゲノミクス・プロテオミクスの新展開

基因组学和蛋白质组学的新进展

• 发表时间: 2004
• 作者: Takagi;D.;今中 忠行 監修
• 通讯作者: 今中 忠行 監修

Ab initio crystal structure determination of spherical viruses that exhibit a centrosymmetric location in the unit cell

从头算晶体结构测定在晶胞中表现出中心对称位置的球形病毒

• 发表时间: 2005
• 期刊: Acta Crystallographica sect. D 61
• 作者: 齊藤修;増保生郎;T.Watanabe;Y.Miyake;Mayumi Hirose;A.Wada;Y.Yamaguchi;Aki Nagata;Y.Nishimura;T.Shimizu;Shigeru Hashimoto;S.Komiya;Tomoyo Takai;C.Kimura-Yoshida;Miho Kihara;Kenji Inaba;Aki Nagata;Junichiro Taka
• 通讯作者: Junichiro Taka

生命秩序を担う生体超分子

负责生命秩序的生物超分子

• 发表时间: 2005
• 作者: 大村敏博

Using cryoloops for X-ray data collection from protein crystals at room temperature : A simple applicable method

使用 Cryoloops 在室温下从蛋白质晶体收集 X 射线数据：一种简单适用的方法

• 发表时间: 2005
• 期刊: Journal of Crystal Growth 251
• 作者: Toyo-oka;T.;et al.;Shu Jie Li
• 通讯作者: Shu Jie Li