ジャーナル: Nat Commun / 年: 2023 タイトル: SARS-CoV-2 nsp3 and nsp4 are minimal constituents of a pore spanning replication organelle. 著者: Liv Zimmermann / Xiaohan Zhao / Jana Makroczyova / Moritz Wachsmuth-Melm / Vibhu Prasad / Zach Hensel / Ralf Bartenschlager / Petr Chlanda / 要旨: Coronavirus replication is associated with the remodeling of cellular membranes, resulting in the formation of double-membrane vesicles (DMVs). A DMV-spanning pore was identified as a putative portal ...Coronavirus replication is associated with the remodeling of cellular membranes, resulting in the formation of double-membrane vesicles (DMVs). A DMV-spanning pore was identified as a putative portal for viral RNA. However, the exact components and the structure of the SARS-CoV-2 DMV pore remain to be determined. Here, we investigate the structure of the DMV pore by in situ cryo-electron tomography combined with subtomogram averaging. We identify non-structural protein (nsp) 3 and 4 as minimal components required for the formation of a DMV-spanning pore, which is dependent on nsp3-4 proteolytic cleavage. In addition, we show that Mac2-Mac3-DPUP-Ubl2 domains are critical for nsp3 oligomerization and crown integrity which influences membrane curvature required for biogenesis of DMVs. Altogether, SARS-CoV-2 nsp3-4 have a dual role by driving the biogenesis of replication organelles and assembly of DMV-spanning pores which we propose here to term replicopores.
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 / 集束イオンビーム - 電流: 0.03 / 集束イオンビーム - 時間: 200 / 集束イオンビーム - 温度: 93 K / 集束イオンビーム - Initial thickness: 1000 / 集束イオンビーム - 最終 厚さ: 200 集束イオンビーム - 詳細: The value given for _em_focused_ion_beam.instrument is Aquilos. This is not in a list of allowed values {'DB235', 'OTHER'} so OTHER is written into the XML file.
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電子顕微鏡法
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フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 平均電子線量: 3.0 e/Å2