ジャーナル: Cell / 年: 2007 タイトル: RF3 induces ribosomal conformational changes responsible for dissociation of class I release factors. 著者: Haixiao Gao / Zhihong Zhou / Urmila Rawat / Chenhui Huang / Lamine Bouakaz / Chernhoe Wang / Zhihong Cheng / Yuying Liu / Andrey Zavialov / Richard Gursky / Suparna Sanyal / Måns Ehrenberg / ...著者: Haixiao Gao / Zhihong Zhou / Urmila Rawat / Chenhui Huang / Lamine Bouakaz / Chernhoe Wang / Zhihong Cheng / Yuying Liu / Andrey Zavialov / Richard Gursky / Suparna Sanyal / Måns Ehrenberg / Joachim Frank / Haiwei Song / 要旨: During translation termination, class II release factor RF3 binds to the ribosome to promote rapid dissociation of a class I release factor (RF) in a GTP-dependent manner. We present the crystal ...During translation termination, class II release factor RF3 binds to the ribosome to promote rapid dissociation of a class I release factor (RF) in a GTP-dependent manner. We present the crystal structure of E. coli RF3*GDP, which has a three-domain architecture strikingly similar to the structure of EF-Tu*GTP. Biochemical data on RF3 mutants show that a surface region involving domains II and III is important for distinct steps in the action cycle of RF3. Furthermore, we present a cryo-electron microscopy (cryo-EM) structure of the posttermination ribosome bound with RF3 in the GTP form. Our data show that RF3*GTP binding induces large conformational changes in the ribosome, which break the interactions of the class I RF with both the decoding center and the GTPase-associated center of the ribosome, apparently leading to the release of the class I RF.
モード: BRIGHT FIELD / 倍率(公称値): 50000 X / 倍率(補正後): 49696 X / 最大 デフォーカス(公称値): 4000 nm / 最小 デフォーカス(公称値): 2000 nm / Cs: 2 mm
試料ホルダ
温度: 93 K / 傾斜角・最大: 0 ° / 傾斜角・最小: 0 °
撮影
電子線照射量: 20 e/Å2 / フィルム・検出器のモデル: KODAK SO-163 FILM
放射
プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M / 散乱光タイプ: x-ray
放射波長
相対比: 1
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解析
EMソフトウェア
ID
名称
カテゴリ
詳細
1
RSRef
モデルフィッティング
2
RSRef
モデルフィッティング
TNT
3
SPIDER
3次元再構成
CTF補正
詳細: CTF correction of 3D map
対称性
点対称性: C1 (非対称)
3次元再構成
手法: Single particle, Reference-based alignment / 解像度: 15.5 Å / 解像度の算出法: FSC 0.5 CUT-OFF / 粒子像の数: 45000 / ピクセルサイズ(公称値): 2.8 Å / ピクセルサイズ(実測値): 2.82 Å / 倍率補正: TMV 詳細: SPIDER package, 0.5 cutoff of FSC, This entry contains CA atoms only 対称性のタイプ: POINT
原子モデル構築
プロトコル: RIGID BODY FIT / 空間: REAL / Target criteria: cross-correlation coefficient 詳細: METHOD--auto REFINEMENT PROTOCOL--Multi-rigid body, real-space refinement