ジャーナル: Cell / 年: 2018 タイトル: In Situ Structure of Neuronal C9orf72 Poly-GA Aggregates Reveals Proteasome Recruitment. 著者: Qiang Guo / Carina Lehmer / Antonio Martínez-Sánchez / Till Rudack / Florian Beck / Hannelore Hartmann / Manuela Pérez-Berlanga / Frédéric Frottin / Mark S Hipp / F Ulrich Hartl / Dieter ...著者: Qiang Guo / Carina Lehmer / Antonio Martínez-Sánchez / Till Rudack / Florian Beck / Hannelore Hartmann / Manuela Pérez-Berlanga / Frédéric Frottin / Mark S Hipp / F Ulrich Hartl / Dieter Edbauer / Wolfgang Baumeister / Rubén Fernández-Busnadiego / 要旨: Protein aggregation and dysfunction of the ubiquitin-proteasome system are hallmarks of many neurodegenerative diseases. Here, we address the elusive link between these phenomena by employing cryo- ...Protein aggregation and dysfunction of the ubiquitin-proteasome system are hallmarks of many neurodegenerative diseases. Here, we address the elusive link between these phenomena by employing cryo-electron tomography to dissect the molecular architecture of protein aggregates within intact neurons at high resolution. We focus on the poly-Gly-Ala (poly-GA) aggregates resulting from aberrant translation of an expanded GGGGCC repeat in C9orf72, the most common genetic cause of amyotrophic lateral sclerosis and frontotemporal dementia. We find that poly-GA aggregates consist of densely packed twisted ribbons that recruit numerous 26S proteasome complexes, while other macromolecules are largely excluded. Proximity to poly-GA ribbons stabilizes a transient substrate-processing conformation of the 26S proteasome, suggesting stalled degradation. Thus, poly-GA aggregates may compromise neuronal proteostasis by driving the accumulation and functional impairment of a large fraction of cellular proteasomes.
A: 12667.681 Å / B: 12667.681 Å / C: 971.28 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
13.680001079914
13.680001079914
13.68
M x/y/z
926
926
71
origin x/y/z
0.000
0.000
0.000
length x/y/z
12667.681
12667.681
971.280
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
11
NC/NR/NS
926
926
71
D min/max/mean
-21541.451
6443.688
0.257
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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 : in situ tomogram from Rat neuron
全体
名称: in situ tomogram from Rat neuron
要素
細胞: in situ tomogram from Rat neuron
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超分子 #1: in situ tomogram from Rat neuron
超分子
名称: in situ tomogram from Rat neuron / タイプ: cell / ID: 1 / 親要素: 0 / 含まれる分子: #1-#32
由来(天然)
生物種: Rattus norvegicus (ドブネズミ)
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実験情報
-
構造解析
手法
クライオ電子顕微鏡法
解析
電子線トモグラフィー法
試料の集合状態
cell
-
試料調製
緩衝液
pH: 7
グリッド
モデル: Quantifoil R2/1 / 材質: GOLD / メッシュ: 200 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 雰囲気: AIR / 詳細: The grid was coated with C prior to use
凍結
凍結剤: ETHANE-PROPANE / チャンバー内湿度: 100 % / チャンバー内温度: 298 K / 装置: FEI VITROBOT MARK IV
詳細
FIB milled rat primary neurons
切片作成
集束イオンビーム - 装置: OTHER / 集束イオンビーム - イオン: OTHER / 集束イオンビーム - 電圧: 30 kV / 集束イオンビーム - 電流: 0.5 nA / 集束イオンビーム - Dose rate: 1 / 集束イオンビーム - 時間: 100 sec. / 集束イオンビーム - 温度: 90 K / 集束イオンビーム - Initial thickness: 1500 / 集束イオンビーム - 最終 厚さ: 200 集束イオンビーム - 詳細: 11nA rough milling. The value given for _emd_sectioning_focused_ion_beam.instrument is FEI Scios. This is not in a list of allowed values set(['DB235', 'OTHER']) so ...集束イオンビーム - 詳細: 11nA rough milling. The value given for _emd_sectioning_focused_ion_beam.instrument is FEI Scios. This is not in a list of allowed values set(['DB235', 'OTHER']) so OTHER is written into the XML file.