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-Structure paper
タイトル | A network of mixed actin polarity in the leading edge of spreading cells. |
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ジャーナル・号・ページ | Commun Biol, Vol. 5, Issue 1, Page 1338, Year 2022 |
掲載日 | 2022年12月7日 |
著者 | Wen-Lu Chung / Matthias Eibauer / Wenhong Li / Rajaa Boujemaa-Paterski / Benjamin Geiger / Ohad Medalia / |
PubMed 要旨 | Physical interactions of cells with the underlying extracellular matrix (ECM) play key roles in multiple cellular processes. The actin cytoskeleton is a central driver and regulator of cellular ...Physical interactions of cells with the underlying extracellular matrix (ECM) play key roles in multiple cellular processes. The actin cytoskeleton is a central driver and regulator of cellular dynamics, that produces membrane-protrusions such as lamellipodia and filopodia. Here, we examined actin organization in expanding lamellipodia during early stages of cell spreading. To gain insight into the 3D actin organization, we plated fibroblasts on galectin-8 coated EM grids, an ECM protein presents in disease states. We then combined cryo-electron tomography with advanced image processing tools for reconstructing the structure of F-actin in the lamellipodia. This approach enabled us to resolve the polarity and orientation of filaments, and the structure of the Arp2/3 complexes associated with F-actin branches. We show that F-actin in lamellipodial protrusions forms a dense network with three distinct sub-domains. One consists primarily of radial filaments, with their barbed ends pointing towards the membrane, the other is enriched with parallel filaments that run between the radial fibers, in addition to an intermediate sub-domain. Surprisingly, a minor, yet significant (~10%) population of actin filaments, are oriented with their barbed-ends towards the cell center. Our results provide structural insights into F-actin assembly and dynamic reorganization in the leading edge of spreading cells. |
リンク | Commun Biol / PubMed:36473943 / PubMed Central |
手法 | EM (トモグラフィー) / EM (サブトモグラム平均) |
解像度 | 14.7 - 25.99 Å |
構造データ | EMDB-15638: Tomogram of lamellipodia EMDB-15644: Tomogram of lamellipodia EMDB-15645: Tomogram of lamellipodia EMDB-15666: Actin filament from lamellipodia of mouse embryonic fibroblasts EMDB-15667: Arp2/3 complex from lamellipodia of mouse embryonic fibroblast |
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