- EMDB-6722: Drosophila full length cryoEM structure of C3PO complex -
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IDまたはキーワード:
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基本情報
登録情報
データベース: EMDB / ID: EMD-6722
タイトル
Drosophila full length cryoEM structure of C3PO complex
マップデータ
cryo-EM structure of full-length C3PO (WT) adopt a closed football-like shape with a hollow interior cavity consisting of four Trax/Translin heterodimers.
ジャーナル: Nucleic Acids Res / 年: 2018 タイトル: Structural insights into Drosophila-C3PO complex assembly and 'Dynamic Side Port' model in substrate entry and release. 著者: Xiaobing Mo / Xia Yang / Yuren Adam Yuan / 要旨: In Drosophila and human, component 3 promoter of RISC (C3PO), a heteromeric complex, enhances RISC assembly and promotes RISC activity. Here, we report crystal structure of full-length Drosophila ...In Drosophila and human, component 3 promoter of RISC (C3PO), a heteromeric complex, enhances RISC assembly and promotes RISC activity. Here, we report crystal structure of full-length Drosophila C3PO (E126Q), an inactive C3PO mutant displaying much weaker RNA binding ability, at 2.1 Å resolution. In addition, we also report the cryo-EM structures of full-length Drosophila C3PO (E126Q), C3PO (WT) and SUMO-C3PO (WT, sumo-TRAX + Translin) particles trapped at different conformations at 12, 19.7 and 12.8 Å resolutions, respectively. Crystal structure of C3PO (E126Q) displays a half-barrel architecture consisting of two Trax/Translin heterodimers, whereas cryo-EM structures of C3PO (E126Q), C3PO (WT) and SUMO-C3PO (WT) adopt a closed football-like shape with a hollow interior cavity. Remarkably, both cryo-EM structures of Drosophila C3PO (E126Q) and Drosophila SUMO-C3PO (WT) particles contain a wide side port (∼25 Å × ∼30 Å versus ∼15 Å × ∼20 Å) for RNA substrate entry and release, formed by a pair of anti-parallel packed long α1 helices of TRAX subunits. Notably, cryo-EM structure of SUMO-C3PO showed that four copies of extra densities belonging to N-terminal SUMO tag are located at the outside shell of SUMO-C3PO particle, which demonstrated that the stoichiometry of TRAX/Translin for the in vitro expressed and assembled full-length Drosophila-SUMO-C3PO particle is 4:4, suggesting Drosophila C3PO is composed by TRAX/translin at a ratio of 4:4. Remarkably, the comparison of the cryo-EM structures suggests that the C3PO side ports regulated by α1 helices of TRAX molecules are highly dynamic. Hence, we propose that C3PO particles could adopt a 'Dynamic Side Port' model to capture/digest nucleic acid duplex substrate and release the digested fragments through the dynamic side ports.
ダウンロード / ファイル: emd_6722.map.gz / 形式: CCP4 / 大きさ: 30.5 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
cryo-EM structure of full-length C3PO (WT) adopt a closed football-like shape with a hollow interior cavity consisting of four Trax/Translin heterodimers.
ボクセルのサイズ
X=Y=Z: 1.14 Å
密度
表面レベル
登録者による: 3.86 / ムービー #1: 3.86
最小 - 最大
-4.7129374 - 10.570582
平均 (標準偏差)
0.15714525 (±1.1022432)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
-100
-100
-100
サイズ
200
200
200
Spacing
200
200
200
セル
A=B=C: 228.0 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
1.14
1.14
1.14
M x/y/z
200
200
200
origin x/y/z
0.000
0.000
0.000
length x/y/z
228.000
228.000
228.000
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
-100
-100
-100
NC/NR/NS
200
200
200
D min/max/mean
-4.713
10.571
0.157
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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 : C3PO
全体
名称: C3PO
要素
複合体: C3PO
複合体: TRAX
複合体: Translin
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超分子 #1: C3PO
超分子
名称: C3PO / タイプ: complex / ID: 1 / 親要素: 0 詳細: In Drosophila, component 3 promoter of RISC (C3PO)is a complex, consisting of 4 Trax/Translin heterodimers.