ジャーナル: Nucleic Acids Res / 年: 2013 タイトル: Promiscuous behaviour of archaeal ribosomal proteins: implications for eukaryotic ribosome evolution. 著者: Jean-Paul Armache / Andreas M Anger / Viter Márquez / Sibylle Franckenberg / Thomas Fröhlich / Elizabeth Villa / Otto Berninghausen / Michael Thomm / Georg J Arnold / Roland Beckmann / Daniel N Wilson / 要旨: In all living cells, protein synthesis occurs on ribonucleoprotein particles called ribosomes. Molecular models have been reported for complete bacterial 70S and eukaryotic 80S ribosomes; however, ...In all living cells, protein synthesis occurs on ribonucleoprotein particles called ribosomes. Molecular models have been reported for complete bacterial 70S and eukaryotic 80S ribosomes; however, only molecular models of large 50S subunits have been reported for archaea. Here, we present a complete molecular model for the Pyrococcus furiosus 70S ribosome based on a 6.6 Å cryo-electron microscopy map. Moreover, we have determined cryo-electron microscopy reconstructions of the Euryarchaeota Methanococcus igneus and Thermococcus kodakaraensis 70S ribosomes and Crenarchaeota Staphylothermus marinus 50S subunit. Examination of these structures reveals a surprising promiscuous behavior of archaeal ribosomal proteins: We observe intersubunit promiscuity of S24e and L8e (L7ae), the latter binding to the head of the small subunit, analogous to S12e in eukaryotes. Moreover, L8e and L14e exhibit intrasubunit promiscuity, being present in two copies per archaeal 50S subunit, with the additional binding site of L14e analogous to the related eukaryotic r-protein L27e. Collectively, these findings suggest insights into the evolution of eukaryotic ribosomal proteins through increased copy number and binding site promiscuity.
タイプ: NEGATIVE 詳細: 20 mM Hepes pH 7.5, 10 mM Mg(OAc)2, 30 mM NH4OAc, 4 mM beta-Mercaptoethanol
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: FEI VITROBOT MARK I / 詳細: Vitrification instrument: Vitrobot 手法: Blot for 10 seconds before plunging, use 2 layers of filter paper
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電子顕微鏡法
顕微鏡
FEI TECNAI 12
日付
2008年8月9日
撮影
カテゴリ: FILM / フィルム・検出器のモデル: FEI EAGLE (4k x 4k) / デジタル化 - スキャナー: OTHER / 実像数: 99 / 平均電子線量: 20 e/Å2 / 詳細: Eagle 4k CCD
電子線
加速電圧: 120 kV / 電子線源: LAB6
電子光学系
倍率(補正後): 90000 / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.0 mm / 倍率(公称値): 90000
試料ステージ
試料ホルダーモデル: GATAN LIQUID NITROGEN
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画像解析
CTF補正
詳細: Wiener filter on 3D volumes
最終 再構成
想定した対称性 - 点群: C1 (非対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 19.0 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: SPIDER / 使用した粒子像数: 10431
Protocol: Rigid-body fit into the density using UCSF Chimera built-in "Fit in Map". Rigid-body fit into the density using UCSF Chimera built-in "Fit in Map"