ジャーナル: Nature / 年: 2008 タイトル: 3.88 A structure of cytoplasmic polyhedrosis virus by cryo-electron microscopy. 著者: Xuekui Yu / Lei Jin / Z Hong Zhou / 要旨: Cytoplasmic polyhedrosis virus (CPV) is unique within the Reoviridae family in having a turreted single-layer capsid contained within polyhedrin inclusion bodies, yet being fully capable of cell ...Cytoplasmic polyhedrosis virus (CPV) is unique within the Reoviridae family in having a turreted single-layer capsid contained within polyhedrin inclusion bodies, yet being fully capable of cell entry and endogenous RNA transcription. Biochemical data have shown that the amino-terminal 79 residues of the CPV turret protein (TP) is sufficient to bring CPV or engineered proteins into the polyhedrin matrix for micro-encapsulation. Here we report the three-dimensional structure of CPV at 3.88 A resolution using single-particle cryo-electron microscopy. Our map clearly shows the turns and deep grooves of alpha-helices, the strand separation in beta-sheets, and densities for loops and many bulky side chains; thus permitting atomic model-building effort from cryo-electron microscopy maps. We observed a helix-to-beta-hairpin conformational change between the two conformational states of the capsid shell protein in the region directly interacting with genomic RNA. We have also discovered a messenger RNA release hole coupled with the mRNA capping machinery unique to CPV. Furthermore, we have identified the polyhedrin-binding domain, a structure that has potential in nanobiotechnology applications.
名称: Bombyx mori cypovirus 1 / タイプ: virus / ID: 1 / Name.synonym: CPV 詳細: CPV is an unenveloped virus wiht a single-shell capsid and diameter of 750 Angstroms. NCBI-ID: 110829 / 生物種: Bombyx mori cypovirus 1 / ウイルスタイプ: VIRION / ウイルス・単離状態: STRAIN / ウイルス・エンベロープ: No / ウイルス・中空状態: No / Syn species name: CPV
宿主
生物種: Bombyx mori (カイコ) / 別称: INVERTEBRATES
ウイルス殻
Shell ID: 1 / 直径: 750 Å / T番号(三角分割数): 1
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実験情報
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構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
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試料調製
緩衝液
pH: 7.4 / 詳細: 10mM PBS
グリッド
詳細: the holes of holey carbon films
凍結
凍結剤: ETHANE / チャンバー内温度: 100 K / 装置: HOMEMADE PLUNGER 詳細: Vitrification instrument: lab-made plunger. Vitrification was carried out at room temperature. CPV were embedded in a thin layer of vitreous ice suspended across the holes of holey carbon ...詳細: Vitrification instrument: lab-made plunger. Vitrification was carried out at room temperature. CPV were embedded in a thin layer of vitreous ice suspended across the holes of holey carbon films for cryoEM imaging. 手法: blot for 3 seconds wiht filter paper before plunging
Focal pairs of micrographs were recorded on 4KX4K charge-coupled device (CCD) camera.
CTF補正
詳細: each particle
最終 再構成
想定した対称性 - 点群: I (正20面体型対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 3.88 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: IMIRS 詳細: Determination of particle orientation and center parameters and subsequent 3D reconstruction were carried out using programs in the IMIRS software package, which are based on Fourier common ...詳細: Determination of particle orientation and center parameters and subsequent 3D reconstruction were carried out using programs in the IMIRS software package, which are based on Fourier common lines and Fourier-Bessel synthesis methods. Prior to the merging of particles for 3D reconstruction, the Fourier transform values of individual images were corrected for the CTF with 15 percent amplitude contrast and a decay factor of 35 sq. Angstroms. 使用した粒子像数: 12814