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基本情報
登録情報 | ![]() | |||||||||
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タイトル | Electron tomogram of ER-nuclear envelope junction of HeLa cell in early telophase | |||||||||
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![]() | Endoplasmic reticulum / Nuclear envelope / Organelle contact site / Membrane junction / CELL CYCLE | |||||||||
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手法 | 電子線トモグラフィー法 / クライオ電子顕微鏡法 / ネガティブ染色法 | |||||||||
![]() | Bragulat-Teixidor H / Otsuka S | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: The endoplasmic reticulum connects to the nucleus by constricted junctions that mature after mitosis. 著者: Helena Bragulat-Teixidor / Keisuke Ishihara / Gréta Martina Szücs / Shotaro Otsuka / ![]() ![]() 要旨: Junctions between the endoplasmic reticulum (ER) and the outer membrane of the nuclear envelope (NE) physically connect both organelles. These ER-NE junctions are essential for supplying the NE with ...Junctions between the endoplasmic reticulum (ER) and the outer membrane of the nuclear envelope (NE) physically connect both organelles. These ER-NE junctions are essential for supplying the NE with lipids and proteins synthesized in the ER. However, little is known about the structure of these ER-NE junctions. Here, we systematically study the ultrastructure of ER-NE junctions in cryo-fixed mammalian cells staged in anaphase, telophase, and interphase by correlating live cell imaging with three-dimensional electron microscopy. Our results show that ER-NE junctions in interphase cells have a pronounced hourglass shape with a constricted neck of 7-20 nm width. This morphology is significantly distinct from that of junctions within the ER network, and their morphology emerges as early as telophase. The highly constricted ER-NE junctions are seen in several mammalian cell types, but not in budding yeast. We speculate that the unique and highly constricted ER-NE junctions are regulated via novel mechanisms that contribute to ER-to-NE lipid and protein traffic in higher eukaryotes. | |||||||||
履歴 |
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構造の表示
添付画像 |
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マップデータ | ![]() | 14.7 GB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 9 KB 9 KB | 表示 表示 | ![]() |
画像 | ![]() | 232.8 KB | ||
Filedesc metadata | ![]() | 3.9 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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リンク
EMDBのページ | ![]() ![]() |
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マップ
ファイル | ![]() | ||||||||||||||||||||
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ボクセルのサイズ | X=Y=Z: 4.51 Å | ||||||||||||||||||||
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対称性 | 空間群: 1 | ||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
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試料の構成要素
-全体 : HeLa Kyoto cell
全体 | 名称: HeLa Kyoto cell |
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要素 |
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-超分子 #1: HeLa Kyoto cell
超分子 | 名称: HeLa Kyoto cell / タイプ: cell / ID: 1 / 親要素: 0 |
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由来(天然) | 生物種: ![]() |
-実験情報
-構造解析
手法 | ネガティブ染色法, クライオ電子顕微鏡法 |
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![]() | 電子線トモグラフィー法 |
試料の集合状態 | cell |
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試料調製
緩衝液 | pH: 7.4 詳細: DMEM without Riboflavin and Phenol Red, containing 10% FBS, 1% Pen/Strep, and 50 nM SiR-DNA |
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染色 | タイプ: NEGATIVE / 材質: Uranyl acetate and lead citrate |
糖包埋 | 材質: Agar 100 Epoxy resin 詳細: Frozen cells were substituted in 0.1% uranyl acetate (UA), 2% Osmium tetroxide and 5% H2O in acetone following this temperature ramp: -90 C to -80 C for 10 hours, -80 C to -30 C for 10 hours, ...詳細: Frozen cells were substituted in 0.1% uranyl acetate (UA), 2% Osmium tetroxide and 5% H2O in acetone following this temperature ramp: -90 C to -80 C for 10 hours, -80 C to -30 C for 10 hours, -30 C for 4 hours, -30 C to 0 C for 6 ours, 0 C to 20 C for 4 hours, 20 C for 5-6 hours. |
凍結 | 凍結剤: NITROGEN |
加圧凍結法 | 装置: OTHER 詳細: High pressure freezing chamber was 1 mm thick, 6.0 mm diameter, with central cavities 5.0 mm x 5.0 mm x 25 um deep. The chamber had been in contact with 1-hexadecene.. The value given for _em_ ...詳細: High pressure freezing chamber was 1 mm thick, 6.0 mm diameter, with central cavities 5.0 mm x 5.0 mm x 25 um deep. The chamber had been in contact with 1-hexadecene.. The value given for _em_high_pressure_freezing.instrument is Leica EM ICE. This is not in a list of allowed values {'LEICA EM PACT', 'LEICA EM PACT2', 'BAL-TEC HPM 010', 'OTHER', 'LEICA EM HPM100', 'EMS-002 RAPID IMMERSION FREEZER'} so OTHER is written into the XML file. |
Cryo protectant | 20% Ficoll-PM400 |
切片作成 | ウルトラミクロトーム - 装置: Leica Ultracut UCT / ウルトラミクロトーム - 温度: 298 K / ウルトラミクロトーム - 最終 厚さ: 250 |
位置合わせマーカー | Manufacturer: Cytodiagnostics / 直径: 15 nm |
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電子顕微鏡法
顕微鏡 | FEI TECNAI F20 |
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撮影 | フィルム・検出器のモデル: FEI EAGLE (4k x 4k) / 平均電子線量: 60.0 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 0.5 µm / 最小 デフォーカス(公称値): 0.2 µm |
実験機器 | ![]() モデル: Tecnai F20 / 画像提供: FEI Company |
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画像解析
最終 再構成 | アルゴリズム: BACK PROJECTION / 使用した粒子像数: 110 |
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