ジャーナル: Structure / 年: 2015 タイトル: Novel Inter-Subunit Contacts in Barley Stripe Mosaic Virus Revealed by Cryo-Electron Microscopy. 著者: Daniel Kofi Clare / Eugenia V Pechnikova / Eugene V Skurat / Valentin V Makarov / Olga S Sokolova / Andrey G Solovyev / Elena V Orlova / 要旨: Barley stripe mosaic virus (BSMV, genus Hordeivirus) is a rod-shaped single-stranded RNA virus similar to viruses of the structurally characterized and well-studied genus Tobamovirus. Here we report ...Barley stripe mosaic virus (BSMV, genus Hordeivirus) is a rod-shaped single-stranded RNA virus similar to viruses of the structurally characterized and well-studied genus Tobamovirus. Here we report the first high-resolution structure of BSMV at 4.1 Å obtained by cryo-electron microscopy. We discovered that BSMV forms two types of virion that differ in the number of coat protein (CP) subunits per turn and interactions between the CP subunits. While BSMV and tobacco mosaic virus CP subunits have a similar fold and interact with RNA using conserved residues, the axial contacts between the CP of these two viral groups are considerably different. BSMV CP subunits lack substantial axial contacts and are held together by a previously unobserved lateral contact formed at the virion surface via an interacting loop, which protrudes from the CP hydrophobic core to the adjacent CP subunit. These data provide an insight into diversity in structural organization of helical viruses.
名称: chimeric Barley Stripe Mosaic Virus wide virion / タイプ: sample / ID: 1000 詳細: The sample is a helical virus with 111 copies of the capsid protein per helical repeat 集合状態: Helical, with 111 copies of the capsid protein per helical repeat Number unique components: 2
分子量
実験値: 22.5 KDa / 理論値: 22.5 KDa
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超分子 #1: Barley stripe mosaic virus
超分子
名称: Barley stripe mosaic virus / タイプ: virus / ID: 1 / Name.synonym: BSMV / 詳細: Has RNA bound / NCBI-ID: 12327 / 生物種: Barley stripe mosaic virus / Sci species strain: ND18 / 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: BSMV
名称: RNA / タイプ: rna / ID: 1 / Name.synonym: RNA / 分類: OTHER / Structure: SINGLE STRANDED / Synthetic?: No
由来(天然)
生物種: Tobacco mosaic virus (ウイルス)
配列
文字列:
GAA
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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
helical array
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試料調製
濃度
3 mg/mL
緩衝液
pH: 7.5 / 詳細: 50mM Tris-HCL, 50mM KCl, 10mM MgCl2
グリッド
詳細: c-flats r2/2 or home made continuous carbon grids
凍結
凍結剤: ETHANE / チャンバー内湿度: 40 % / チャンバー内温度: 100 K / 装置: HOMEMADE PLUNGER 手法: 3.5ul of sample was added to continuous carbon coated or c-flat grids, blotted for 2 seconds and then plunged in liquid ethane
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電子顕微鏡法
顕微鏡
FEI POLARA 300
温度
最低: 80 K
アライメント法
Legacy - 非点収差: Objective lens astigmatism was corrected at 150,000 times magnification
日付
2012年6月1日
撮影
カテゴリ: CCD フィルム・検出器のモデル: GATAN ULTRASCAN 4000 (4k x 4k) デジタル化 - サンプリング間隔: 15 µm / 実像数: 297 / 平均電子線量: 25 e/Å2 / 詳細: each image was a 1 second low dose exposure / ビット/ピクセル: 16
BPRP was used for reconstruction. The particles were selected using the helical option in boxer. Helical symmetry was imposed, with 111 subunits in the 5 turns forming the helical repeat.
CTF補正
詳細: phase flipped each particle
最終 再構成
想定した対称性 - らせんパラメータ - Δz: 1.182 Å 想定した対称性 - らせんパラメータ - ΔΦ: 16.218 ° 想定した対称性 - らせんパラメータ - 軸対称性: C1 (非対称) アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 4.1 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: eman, Imagic, spider / 詳細: Final maps calculated with BPRP / 使用した粒子像数: 2077
最終 角度割当
詳細: 1 degree steps from 80-100 degrees in beta and from 0-16.216 degrees in gamma