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基本情報
登録情報 | データベース: EMDB / ID: EMD-2369 | |||||||||
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タイトル | The Respiratory Syncytial Virus nucleoprotein-RNA complex forms a left-handed helical nucleocapsid. | |||||||||
![]() | Sub-tomogram average of purified RSV nucleocapsid | |||||||||
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![]() | Respiratory Syncytial Virus / nucleocapsid / cryo-electron tomography / sub-tomogram averaging | |||||||||
機能・相同性 | ![]() Respiratory syncytial virus genome transcription / symbiont-mediated suppression of host PKR/eIFalpha signaling / Translation of respiratory syncytial virus mRNAs / protein serine/threonine kinase inhibitor activity / Respiratory syncytial virus genome replication / helical viral capsid / RSV-host interactions / Maturation of hRSV A proteins / Assembly and release of respiratory syncytial virus (RSV) virions / Respiratory syncytial virus (RSV) attachment and entry ...Respiratory syncytial virus genome transcription / symbiont-mediated suppression of host PKR/eIFalpha signaling / Translation of respiratory syncytial virus mRNAs / protein serine/threonine kinase inhibitor activity / Respiratory syncytial virus genome replication / helical viral capsid / RSV-host interactions / Maturation of hRSV A proteins / Assembly and release of respiratory syncytial virus (RSV) virions / Respiratory syncytial virus (RSV) attachment and entry / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of MDA-5 activity / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of MAVS activity / PKR-mediated signaling / Evasion by RSV of host interferon responses / viral capsid / viral nucleocapsid / host cell cytoplasm / symbiont-mediated suppression of host type I interferon-mediated signaling pathway / ribonucleoprotein complex / virus-mediated perturbation of host defense response / RNA binding 類似検索 - 分子機能 | |||||||||
生物種 | ![]() | |||||||||
手法 | サブトモグラム平均法 / ネガティブ染色法 | |||||||||
![]() | Bakker SE / Duquerroy S / Galloux M / Loney C / Conner E / Eleouet JF / Rey FA / Bhella D | |||||||||
![]() | ![]() タイトル: The respiratory syncytial virus nucleoprotein-RNA complex forms a left-handed helical nucleocapsid. 著者: Saskia E Bakker / Stéphane Duquerroy / Marie Galloux / Colin Loney / Edward Conner / Jean-François Eléouët / Félix A Rey / David Bhella / ![]() ![]() 要旨: Respiratory syncytial virus (RSV) is an important human pathogen. Its nucleocapsid (NC), which comprises the negative sense RNA viral genome coated by the viral nucleoprotein N, is a critical ...Respiratory syncytial virus (RSV) is an important human pathogen. Its nucleocapsid (NC), which comprises the negative sense RNA viral genome coated by the viral nucleoprotein N, is a critical assembly that serves as template for both mRNA synthesis and genome replication. We have previously described the X-ray structure of an NC-like structure: a decameric ring formed of N-RNA that mimics one turn of the helical NC. In the absence of experimental data we had hypothesized that the NC helix would be right-handed, as the N-N contacts in the ring appeared to more easily adapt to that conformation. We now unambiguously show that the RSV NC is a left-handed helix. We further show that the contacts in the ring can be distorted to maintain key N-N-protein interactions in a left-handed helix, and discuss the implications of the resulting atomic model of the helical NC for viral replication and transcription. | |||||||||
履歴 |
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構造の表示
ムービー |
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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マップデータ | ![]() | 217.8 KB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 11.4 KB 11.4 KB | 表示 表示 | ![]() |
画像 | ![]() | 47.6 KB | ||
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-検証レポート
文書・要旨 | ![]() | 200.4 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 199.5 KB | 表示 | |
XML形式データ | ![]() | 4.8 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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リンク
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マップ
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注釈 | Sub-tomogram average of purified RSV nucleocapsid | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 6.5 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
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試料の構成要素
-全体 : Purified recombinant RSV nucleocapsids.
全体 | 名称: Purified recombinant RSV nucleocapsids. |
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要素 |
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-超分子 #1000: Purified recombinant RSV nucleocapsids.
超分子 | 名称: Purified recombinant RSV nucleocapsids. / タイプ: sample / ID: 1000 / 集合状態: variable helices / Number unique components: 1 |
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-超分子 #1: Respiratory syncytial virus
超分子 | 名称: Respiratory syncytial virus / タイプ: virus / ID: 1 / Name.synonym: RSV / 詳細: Purified recombinant nucleocapsid / NCBI-ID: 12814 / 生物種: Respiratory syncytial virus / ウイルスタイプ: OTHER / ウイルス・単離状態: OTHER / ウイルス・エンベロープ: No / ウイルス・中空状態: No / Syn species name: RSV |
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宿主 | 生物種: ![]() |
Host system | 生物種: ![]() ![]() 組換細胞: Sf21 / 組換プラスミド: Rbac69 |
-実験情報
-構造解析
手法 | ネガティブ染色法 |
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![]() | サブトモグラム平均法 |
試料の集合状態 | filament |
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試料調製
緩衝液 | pH: 7.4 / 詳細: 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 10 mM EDTA |
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染色 | タイプ: NEGATIVE 詳細: Protein preparations were mixed with 10 nm colloidal gold and loaded onto freshly glow-discharged perforated C-flat carbon films (CF-22-2C, Protochips Inc.). Grids were then washed in 0.1% ...詳細: Protein preparations were mixed with 10 nm colloidal gold and loaded onto freshly glow-discharged perforated C-flat carbon films (CF-22-2C, Protochips Inc.). Grids were then washed in 0.1% trehalose, and stained using 5% ammonium molybdate (pH 7.4) and 0.1% trehalose, drained and allowed to dry in air |
グリッド | 詳細: Glow-discharged perforated C-flat carbon films (CF-22-2C, Protochips Inc.) |
凍結 | 凍結剤: NONE / 装置: OTHER |
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電子顕微鏡法
顕微鏡 | JEOL 2200FSC |
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温度 | 最低: 97 K / 最高: 293 K |
アライメント法 | Legacy - 非点収差: Corrected at 100k times mag in digital micrograph |
特殊光学系 | エネルギーフィルター - 名称: Jeol エネルギーフィルター - エネルギー下限: 0.0 eV エネルギーフィルター - エネルギー上限: 30.0 eV |
詳細 | Room temperature stained grids imaged at LN2 temperatures. |
日付 | 2010年7月15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) 実像数: 9 / 平均電子線量: 131 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.0 mm / 最大 デフォーカス(公称値): 4.0 µm / 最小 デフォーカス(公称値): 2.0 µm / 倍率(公称値): 40000 |
試料ステージ | 試料ホルダーモデル: GATAN LIQUID NITROGEN / Tilt series - Axis1 - Min angle: -60 ° / Tilt series - Axis1 - Max angle: 60 ° |
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画像解析
詳細 | Subtomograms were manually picked. A starting model was obtained by manually aligning 100 subtomograms, all subtomograms were aligned to this starting model. To mitigate missing wedge artefacts, the orientations around the helical axis were randomised before further alignment. |
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最終 再構成 | アルゴリズム: OTHER / ソフトウェア - 名称: Etomo, Dynamo / 詳細: Subtomograms were picked from nine tomograms. / 使用したサブトモグラム数: 911 |
最終 3次元分類 | クラス数: 1 |