Many of the more than 20 mammalian proteins with N-BAR domains1,2 control cell architecture3 and endocytosis4,5 by associating with curved sections of the plasma membrane6. It is not well understood whether N-BAR proteins are recruited directly by processes that mechanically curve the plasma membrane or indirectly by plasma-membrane-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 plasma membrane 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 plasma membrane. Markedly, live-cell imaging in adherent cells showed selective recruitment of full-length N-BAR proteins and isolated N-BAR domains to plasma membrane 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 plasma membrane sites during local lamellipodia retraction in the front of migrating cells. Recruitment required myosin-II-generated force applied to plasma-membrane-connected actin cables. Together, our results show that N-BAR domains can be directly recruited to the plasma membrane by external push or internal pull forces that locally curve the plasma membrane.
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The authors thank the members of the Meyer laboratory for comments and discussion. M.G. was supported by the Swiss National Science Foundation (No. PBBSP3-123159), and a Novartis Jubilaeumsstiftung and Stanford Deans Postdoctoral Fellowship. S.J. was supported by a Korea Foundation for Advanced Studies graduate fellowship. Y.C. acknowledges the partial support from a DOE-EFRC at Stanford: Center on Nanostructuring for Efficient Energy Conversion (No. DE-SC0001060). T.M. acknowledges financial support from the National Institutes of Health, MH064801, MH095087 and GM063702.
The authors declare no competing financial interests.
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Galic, M., Jeong, S., Tsai, FC. et al. External push and internal pull forces recruit curvature-sensing N-BAR domain proteins to the plasma membrane. Nat Cell Biol 14, 874–881 (2012). https://doi.org/10.1038/ncb2533
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