National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R35GM118149
米国
Human Frontier Science Program (HFSP)
LT000234/2018-L
フランス
引用
ジャーナル: Dev Cell / 年: 2022 タイトル: Mechanistic insights into actin force generation during vesicle formation from cryo-electron tomography. 著者: Daniel Serwas / Matthew Akamatsu / Amir Moayed / Karthik Vegesna / Ritvik Vasan / Jennifer M Hill / Johannes Schöneberg / Karen M Davies / Padmini Rangamani / David G Drubin / 要旨: Actin assembly provides force for a multitude of cellular processes. Compared to actin-assembly-based force production during cell migration, relatively little is understood about how actin assembly ...Actin assembly provides force for a multitude of cellular processes. Compared to actin-assembly-based force production during cell migration, relatively little is understood about how actin assembly generates pulling forces for vesicle formation. Here, cryo-electron tomography identified actin filament number, organization, and orientation during clathrin-mediated endocytosis in human SK-MEL-2 cells, showing that force generation is robust despite variance in network organization. Actin dynamics simulations incorporating a measured branch angle indicate that sufficient force to drive membrane internalization is generated through polymerization and that assembly is triggered from ∼4 founding "mother" filaments, consistent with tomography data. Hip1R actin filament anchoring points are present along the entire endocytic invagination, where simulations show that it is key to pulling force generation, and along the neck, where it targets filament growth and makes internalization more robust. Actin organization described here allowed direct translation of structure to mechanism with broad implications for other actin-driven processes.
想定した対称性 - 点群: C3 (3回回転対称) / アルゴリズム: BACK PROJECTION / 解像度のタイプ: BY AUTHOR / 解像度: 27.0 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア: (名称: IMOD, PEET) 詳細: Half maps were generated using the fnHalfVolume function of calcFSC in PEET rather than "gold standard" FSC. 使用したサブトモグラム数: 472