ジャーナル: Nat Commun / 年: 2017 タイトル: Ubiquitination of stalled ribosome triggers ribosome-associated quality control. 著者: Yoshitaka Matsuo / Ken Ikeuchi / Yasushi Saeki / Shintaro Iwasaki / Christian Schmidt / Tsuyoshi Udagawa / Fumiya Sato / Hikaru Tsuchiya / Thomas Becker / Keiji Tanaka / Nicholas T Ingolia / ...著者: Yoshitaka Matsuo / Ken Ikeuchi / Yasushi Saeki / Shintaro Iwasaki / Christian Schmidt / Tsuyoshi Udagawa / Fumiya Sato / Hikaru Tsuchiya / Thomas Becker / Keiji Tanaka / Nicholas T Ingolia / Roland Beckmann / Toshifumi Inada / 要旨: Translation arrest by polybasic sequences induces ribosome stalling, and the arrest product is degraded by the ribosome-mediated quality control (RQC) system. Here we report that ubiquitination of ...Translation arrest by polybasic sequences induces ribosome stalling, and the arrest product is degraded by the ribosome-mediated quality control (RQC) system. Here we report that ubiquitination of the 40S ribosomal protein uS10 by the E3 ubiquitin ligase Hel2 (or RQT1) is required for RQC. We identify a RQC-trigger (RQT) subcomplex composed of the RNA helicase-family protein Slh1/Rqt2, the ubiquitin-binding protein Cue3/Rqt3, and yKR023W/Rqt4 that is required for RQC. The defects in RQC of the RQT mutants correlate with sensitivity to anisomycin, which stalls ribosome at the rotated form. Cryo-electron microscopy analysis reveals that Hel2-bound ribosome are dominantly the rotated form with hybrid tRNAs. Ribosome profiling reveals that ribosomes stalled at the rotated state with specific pairs of codons at P-A sites serve as RQC substrates. Rqt1 specifically ubiquitinates these arrested ribosomes to target them to the RQT complex, allowing subsequent RQC reactions including dissociation of the stalled ribosome into subunits.Several protein quality control mechanisms are in place to trigger the rapid degradation of aberrant polypeptides and mRNAs. Here the authors describe a mechanism of ribosome-mediated quality control that involves the ubiquitination of ribosomal proteins by the E3 ubiquitin ligase Hel2/RQT1.
ダウンロード / ファイル: emd_3736.map.gz / 形式: CCP4 / 大きさ: 8.4 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Class 1 of rotated ribosomes (rotated 2 state) with A/P- and P/E-site tRNAs of the Rqt1-FTP pulldown
ボクセルのサイズ
X=Y=Z: 3.252 Å
密度
表面レベル
登録者による: 0.027 / ムービー #1: 0.027
最小 - 最大
-0.090526015 - 0.24486277
平均 (標準偏差)
-0.00048580748 (±0.02409707)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
130
130
130
Spacing
130
130
130
セル
A=B=C: 422.76 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
3.252
3.252
3.252
M x/y/z
130
130
130
origin x/y/z
0.000
0.000
0.000
length x/y/z
422.760
422.760
422.760
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
130
130
130
D min/max/mean
-0.091
0.245
-0.000
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添付データ
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試料の構成要素
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全体 : Class 1 of rotated ribosomes (rotated 2 state) with A/P- and P/E-...
全体
名称: Class 1 of rotated ribosomes (rotated 2 state) with A/P- and P/E-site tRNAs of the Rqt1-FTP pulldown
要素
複合体: Class 1 of rotated ribosomes (rotated 2 state) with A/P- and P/E-site tRNAs of the Rqt1-FTP pulldown
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超分子 #1: Class 1 of rotated ribosomes (rotated 2 state) with A/P- and P/E-...
超分子
名称: Class 1 of rotated ribosomes (rotated 2 state) with A/P- and P/E-site tRNAs of the Rqt1-FTP pulldown タイプ: complex / ID: 1 / 親要素: 0 詳細: Fourth class from classification for dataset in Frealign
由来(天然)
生物種: Saccharomyces cerevisiae (パン酵母)
分子量
理論値: 3.6 MDa
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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
-
試料調製
緩衝液
pH: 7.5
凍結
凍結剤: ETHANE
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
撮影
フィルム・検出器のモデル: FEI FALCON II (4k x 4k) 平均電子線量: 28.0 e/Å2
電子線
加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN
電子光学系
照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm
実験機器
モデル: Titan Krios / 画像提供: FEI Company
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画像解析
最終 再構成
解像度のタイプ: BY AUTHOR / 解像度: 7.0 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 38399