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External push and internal pull forces recruit curvature sensing N-BAR domain proteins to the plasma membrane

机译:外部推和内部拉力招募曲率感测的N- BaR域蛋白至质膜

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摘要

Many of the more than 20 mammalian proteins with N-BAR domains- control cell architecture and endocytosis- by associating with curved sections of the plasma membrane (PM). It is not well understood whether N-BAR proteins are recruited directly by processes that mechanically curve the PM or indirectly by PM-associated adaptor proteins that recruit proteins with N-BAR domains that then induce membrane curvature. Here, we show that externally-induced inward deformation of the PM by cone-shaped nanostructures (Nanocones) and internally-induced inward deformation by contracting actin cables both trigger recruitment of isolated N-BAR domains to the curved PM. Markedly, live-cell imaging in adherent cells showed selective recruitment of full length N-BAR proteins and isolated N-BAR domains to PM sub-regions above Nanocone stripes. Electron microscopy confirmed that N-BAR domains are recruited to local membrane sites curved by Nanocones. We further showed that N-BAR domains are periodically recruited to curved PM sites during local lamellipodia retraction in the front of migrating cells. Recruitment required Myosin II-generated force applied to PM connected actin cables. Together, our study shows that N-BAR domains can be directly recruited to the PM by external push or internal pull forces that locally curve the PM.
机译:具有N-Bar结构域的20多种哺乳动物蛋白质 - 控制单元结构 和内吞作用,通过与质膜的弯曲部分相关联(PM) 。还不明白N-Bar蛋白是否直接通过机械曲线曲线的过程或间接地通过募集蛋白质的PM相关的衔接蛋白,然后诱导膜曲率的N-BAR结构域。在这里,我们表明通过锥形纳米结构(纳米载体)的外部引起的PM向内变形以及通过收缩肌动蛋白电缆的内部诱导的内心变形均触发分离的N-BAR域与弯曲PM的触发募集。明显地,粘附细胞中的活细胞成像显示出全长N-BAR蛋白的选择性募集和分离的N-BAR结构域至纳米条纹上方的PM子区域。电子显微镜证实,N-Bar结构域被征收到纳米核糖弯曲的局部膜位点。我们进一步表明,在迁移细胞前面的局部薄片缩回期间,周期性地征收N-BAR域在局部次数缩回期间弯曲PM位点。招聘需要肌蛋白II-产生的力施加到PM连接的肌动蛋白电缆。我们的研究一起表明,N-Bar域可以通过局部推绕PM的外部推动或内部拉力直接招募到PM。

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