Direct Arp2/3-vinculin binding is required for pseudopod extension, but only on compliant substrates and in 3D

IF 4.6 2区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

iScience Pub Date : 2025-05-09 DOI:10.1016/j.isci.2025.112623

Tadamoto Isogai , Kevin M. Dean , Philippe Roudot , Evgenia V. Azarova , Kushal Bhatt , Meghan K. Driscoll , Shaina P. Royer , Nikhil Mittal , Bo-Jui Chang , Sangyoon J. Han , Reto Fiolka , Gaudenz Danuser

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引用次数: 0

Abstract

A critical step in cell morphogenesis is the extension of actin-dense pseudopods, controlled by actin-binding proteins (ABPs). While this process is well-understood on glass coverslips, it is less so in compliant three-dimensional environments. Here, we knocked out a series of ABPs in osteosarcoma cells and evaluated their effect on pseudopod extension on glass surfaces (2D) and in collagen gels (3D). Cells lacking the longest Arp3 gene variant, or with attenuated Arp2/3 activity, had the strongest reduction in pseudopod formation between 2D and 3D. This was largely due to reduced activity of the hybrid Arp2/3-vinculin complex, which was dispensable on glass. Our data suggests that concurrent formation of actin branches and nascent adhesions, supported by Arp2/3-vinculin interactions, is essential to form mechanically stable links between fibrous extracellular matrix and actin in 3D. This highlights how experiments on stiff, planar substrates may conceal actin architectural features that are essential for morphogenesis in 3D.

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假足延伸需要直接结合arp2 /3-血管蛋白，但仅在柔顺的底物上和3D中

细胞形态发生的一个关键步骤是肌动蛋白密集伪足的延伸，由肌动蛋白结合蛋白（ABPs）控制。虽然这个过程在玻璃盖上很容易理解，但在兼容的三维环境中就不那么容易理解了。在这里，我们敲除骨肉瘤细胞中的一系列ABPs，并评估它们对玻璃表面（2D）和胶原凝胶（3D）假足延伸的影响。缺少最长Arp3基因变体或Arp3 /3活性减弱的细胞，在2D和3D之间假足形成的减少幅度最大。这主要是由于杂交arp2 /3-血管蛋白复合物的活性降低，而这在玻璃上是必不可少的。我们的数据表明，在arp2 /3-血管蛋白相互作用的支持下，肌动蛋白分支和新生粘连的同时形成对于纤维细胞外基质和肌动蛋白在3D中形成机械稳定的连接至关重要。这突出了在坚硬的平面基底上的实验如何隐藏肌动蛋白的结构特征，这些特征对三维形态发生至关重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

iScience Multidisciplinary-Multidisciplinary

CiteScore

7.20

自引率

1.70%

发文量

1972

审稿时长

6 weeks

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