National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R35GM142553
米国
National Institutes of Health/National Institute of Diabetes and Digestive and Kidney Disease (NIH/NIDDK)
R01DK125263
米国
National Institutes of Health/National Cancer Institute (NIH/NCI)
R37CA248565
米国
Jane Coffin Childs (JCC) Fund
米国
Swiss National Science Foundation
P2BSP3_188112
スイス
Swiss National Science Foundation
P400PB_199252
スイス
引用
ジャーナル: EMBO J / 年: 2024 タイトル: In situ architecture of Opa1-dependent mitochondrial cristae remodeling. 著者: Michelle Y Fry / Paula P Navarro / Pusparanee Hakim / Virly Y Ananda / Xingping Qin / Juan C Landoni / Sneha Rath / Zintis Inde / Camila Makhlouta Lugo / Bridget E Luce / Yifan Ge / Julie L ...著者: Michelle Y Fry / Paula P Navarro / Pusparanee Hakim / Virly Y Ananda / Xingping Qin / Juan C Landoni / Sneha Rath / Zintis Inde / Camila Makhlouta Lugo / Bridget E Luce / Yifan Ge / Julie L McDonald / Ilzat Ali / Leillani L Ha / Benjamin P Kleinstiver / David C Chan / Kristopher A Sarosiek / Luke H Chao / 要旨: Cristae membrane state plays a central role in regulating mitochondrial function and cellular metabolism. The protein Optic atrophy 1 (Opa1) is an important crista remodeler that exists as two forms ...Cristae membrane state plays a central role in regulating mitochondrial function and cellular metabolism. The protein Optic atrophy 1 (Opa1) is an important crista remodeler that exists as two forms in the mitochondrion, a membrane-anchored long form (l-Opa1) and a processed short form (s-Opa1). The mechanisms for how Opa1 influences cristae shape have remained unclear due to lack of native three-dimensional views of cristae. We perform in situ cryo-electron tomography of cryo-focused ion beam milled mouse embryonic fibroblasts with defined Opa1 states to understand how each form of Opa1 influences cristae architecture. In our tomograms, we observe a variety of cristae shapes with distinct trends dependent on s-Opa1:l-Opa1 balance. Increased l-Opa1 levels promote cristae stacking and elongated mitochondria, while increased s-Opa1 levels correlated with irregular cristae packing and round mitochondria shape. Functional assays indicate a role for l-Opa1 in wild-type apoptotic and calcium handling responses, and show a compromised respiratory function under Opa1 imbalance. In summary, we provide three-dimensional visualization of cristae architecture to reveal relationships between mitochondrial ultrastructure and cellular function dependent on Opa1-mediated membrane remodeling.
ダウンロード / ファイル: emd_43061.map.gz / 形式: CCP4 / 大きさ: 350.9 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
ボクセルのサイズ
X=Y=Z: 22.06 Å
密度
最小 - 最大
-16.908138000000001 - 61.994143999999999
平均 (標準偏差)
12.8177805 (±1.5852029)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
-104
104
-125
サイズ
720
511
250
Spacing
511
720
250
セル
A: 11272.66 Å / B: 15883.199 Å / C: 5515.0 Å α=β=γ: 90.0 °
-
添付データ
-
試料の構成要素
-
全体 : mitochondria in mouse embryonic fibroblasts with deletion of OPA1
全体
名称: mitochondria in mouse embryonic fibroblasts with deletion of OPA1
要素
細胞: mitochondria in mouse embryonic fibroblasts with deletion of OPA1
-
超分子 #1: mitochondria in mouse embryonic fibroblasts with deletion of OPA1
超分子
名称: mitochondria in mouse embryonic fibroblasts with deletion of OPA1 タイプ: cell / ID: 1 / 親要素: 0
由来(天然)
生物種: Mus musculus (ハツカネズミ)
-
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: 7.5
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277.15 K / 装置: FEI VITROBOT MARK IV
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: 5 / 集束イオンビーム - 電流: 0.01 / 集束イオンビーム - 時間: 600 / 集束イオンビーム - 温度: 88 K / 集束イオンビーム - Initial thickness: 1000 / 集束イオンビーム - 最終 厚さ: 150 集束イオンビーム - 詳細: The value given for _em_focused_ion_beam.instrument is TFS Aquilos2. This is not in a list of allowed values {'DB235', 'OTHER'} so OTHER is written into the XML file.
-
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: GATAN K3 BIOQUANTUM (6k x 4k) 平均電子線量: 3.35 e/Å2