- EMDB-16720: Tomogram of an induced protrusion of a Drosophila S2 cell with fi... -
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基本情報
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データベース: EMDB / ID: EMD-16720
タイトル
Tomogram of an induced protrusion of a Drosophila S2 cell with filaments inside the microtubule lumen.
マップデータ
Deconvolved tomogram (binned by four) of an induced protrusion from Drosophila S2 cells with multiple filaments inside the microtubule lumen. From Dataset 7.
試料
細胞: Tomogram of an induced protrusion of a Drosophila S2 cell with filaments inside the microtubule lumen
ジャーナル: bioRxiv / 年: 2023 タイトル: CryoET shows cofilactin filaments inside the microtubule lumen. 著者: Camilla Ventura Santos / Stephen L Rogers / Andrew P Carter / 要旨: Cytoplasmic microtubules are tubular polymers that can harbor small proteins or filaments inside their lumen. The identity of these objects and what causes their accumulation has not been ...Cytoplasmic microtubules are tubular polymers that can harbor small proteins or filaments inside their lumen. The identity of these objects and what causes their accumulation has not been conclusively established. Here, we used cryogenic electron tomography (cryoET) of S2 cell protrusions and found filaments inside the microtubule lumen, which resemble those reported recently in human HAP1 cells. The frequency of these filaments increased upon inhibition of the sarco/endoplasmic reticulum Ca ATPase (SERCA) with the small-molecule drug thapsigargin. Subtomogram averaging showed that the luminal filaments adopt a helical structure reminiscent of cofilin-bound actin (cofilactin). Consistent with this, cofilin was activated in cells under the same conditions that increased luminal filament occurrence. Furthermore, RNAi knock-down of cofilin reduced the frequency of luminal filaments with cofilactin morphology. These results suggest that cofilin activation stimulates its accumulation on actin filaments inside the microtubule lumen.
ダウンロード / ファイル: emd_16720.map.gz / 形式: CCP4 / 大きさ: 2.1 GB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Deconvolved tomogram (binned by four) of an induced protrusion from Drosophila S2 cells with multiple filaments inside the microtubule lumen. From Dataset 7.
A: 10874.08 Å / B: 15321.601 Å / C: 3979.36 Å α=β=γ: 90.0 °
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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 : Tomogram of an induced protrusion of a Drosophila S2 cell with fi...
全体
名称: Tomogram of an induced protrusion of a Drosophila S2 cell with filaments inside the microtubule lumen
要素
細胞: Tomogram of an induced protrusion of a Drosophila S2 cell with filaments inside the microtubule lumen
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超分子 #1: Tomogram of an induced protrusion of a Drosophila S2 cell with fi...
超分子
名称: Tomogram of an induced protrusion of a Drosophila S2 cell with filaments inside the microtubule lumen タイプ: cell / ID: 1 / 親要素: 0 詳細: Cells were treated with 2.5 uM Cytochalasin D for 4 hours and with 2 uM thapsigargin for 5 hours before vitrification. Example tomogram from dataset 7.
モデル: Quantifoil / 材質: GOLD / メッシュ: 200 詳細: Grids were glow discharged for 30s at 20mA and then coated with 0.25 ug/mL Concanavalin A for at least 2 hours at 37 degrees.
凍結
凍結剤: ETHANE / チャンバー内湿度: 95 % / チャンバー内温度: 298.15 K
詳細
Cells were treated with 2.5 uM Cytochalasin D for 4 hours and with 2 uM thapsigargin for 5 hours before vitrification.
フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 平均電子線量: 3.0 e/Å2 詳細: Data was collected at a pixel size of 2.659 A/pixel with a total dose of 121.54 A/e^2.