TY - JOUR
T1 - Ezrin enrichment on curved membranes requires a specific conformation or interaction with a curvature-sensitive partner
AU - Tsai, Feng Ching
AU - Bertin, Aurelie
AU - Bousquet, Hugo
AU - Manzi, John
AU - Senju, Yosuke
AU - Tsai, Meng Chen
AU - Picas, Laura
AU - Miserey-Lenkei, Stephanie
AU - Lappalainen, Pekka
AU - Lemichez, Emmanuel
AU - Coudrier, Evelyne
AU - Bassereau, Patricia
N1 - Funding Information:
We thank B Goud for insightful discussions, C Le Clainche (Institute for Integrative Biology of the Cell, Gif-sur-Yvette, France) and J Pernier for providing actin and advice on actin reconstitution, F Brochard for helping on analysis of the GUV-tethering assay, F Di Federico for handling plasmids, C Prévost for her help for the optical tweezers setup and data analysis, A Di Cicco and D Levy for EM image acquisition and galcer tube preparation, N de Franceschi for helping on the FACS experiments, M Henderson for carefully reading of the manuscript. The authors greatly acknowledge the Cell and Tissue Imaging (PICT-IBiSA), Institut Curie, member of the French National Research Infrastructure France-BioImaging (ANR10-INBS-04). This work was supported by Institut Curie, Centre National de la Recherche Scientifique (CNRS), the Agence Nationale pour la Recherche (grant ANR-15-CE18-0016 to F-CT, M-CT, EL and PB), the Investments for the Future’ LABEX SIGNALIFE (grant ANR-11-LABX-0028–01 for M-CT and EL), the European Research Council (EC and PB partners of the advanced grant, project 339847) and the Human Frontier Science Program Organization (RGP0005/2016 to YS, PL, F-CT, EC and PB). EC and PB groups belong to the CNRS consortium CellTiss, to the Labex CelTisPhyBio (ANR-11-LABX0038) and to Paris Sciences et Lettres (ANR-10-IDEX-0001–02). F-C Tsai was funded by the EMBO Long-Term fellowship (ALTF 1527–2014) and Marie Curie actions (H2020-MSCA-IF-2014, project membrane-ezrin-actin).
Funding Information:
We thank B Goud for insightful discussions, C Le Clainche (Institute for Integrative Biology of the Cell, Gif-sur-Yvette, France) and J Pernier for providing actin and advice on actin reconstitution, F Brochard for helping on analysis of the GUV-tethering assay, F Di Federico for handling plasmids, C Prévost for her help for the optical tweezers setup and data analysis, A Di Cicco and D Levy for EM image acquisition and galcer tube preparation, N de Franceschi for helping on the FACS experiments, M Henderson for carefully reading of the manuscript. The authors greatly acknowledge the Cell and Tissue Imaging (PICT-IBiSA), Institut Curie, member of the French National Research Infrastructure France-BioImaging (ANR10-INBS-04). This work was supported by Institut Curie, Centre National de la Recherche Scientifique (CNRS), the Agence Nationale pour la Recherche (grant ANR-15-CE18-0016 to F-CT, M-CT, EL and PB), the Investments for the Future’ LABEX SIGNALIFE (grant ANR-11-LABX-0028–01 for M-CT and EL), the European Research Council (EC and PB partners of the advanced grant, project 339847) and the Human Frontier Science Program Organization (RGP0005/2016 to YS, PL, F-CT, EC and PB). EC and PB groups belong to the CNRS consortium CellTiss, to the Labex CelTisPhyBio (ANR-11-LABX0038) and to Paris Sciences et Lettres (ANR-10-IDEX-0001–02). F-C Tsai was funded by the EMBO Long-Term fellowship (ALTF 1527–2014) and Marie Curie actions (H2020-MSCA-IF-2014, project membrane-ezrin-actin). European Molecular Biology Organization ALTF 1527-2014 Feng-Ching Tsai H2020 Marie Skłodowska-Curie Actions H2020-MSCA-IF-2014 Feng-Ching Tsai Human Frontier Science Program RGP0005/2016 Feng-Ching Tsai Yosuke Senju Pekka Lappalainen Evelyne Coudrier Patricia Bassereau Agence Nationale de la Recherche ANR-15-CE18-0016 Feng-Ching Tsai Meng-Chen Tsai Emmanuel Lemichez Patricia Bassereau Investments for the Future LA-BEX SIGNALIFE ANR-11-LABX-0028-01 Meng-Chen Tsai Emmanuel Lemichez H2020 European Research Council 339847 Stephanie Miserey-Lenkei Evelyne Coudrier Patricia Bassereau The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Publisher Copyright:
© Tsai et al.
PY - 2018/9
Y1 - 2018/9
N2 - One challenge in cell biology is to decipher the biophysical mechanisms governing protein enrichment on curved membranes and the resulting membrane deformation. The ERM protein ezrin is abundant and associated with cellular membranes that are flat, positively or negatively curved. Using in vitro and cell biology approaches, we assess mechanisms of ezrin’s enrichment on curved membranes. We evidence that wild-type ezrin (ezrinWT) and its phosphomimetic mutant T567D (ezrinTD) do not deform membranes but self-assemble anti-parallelly, zipping adjacent membranes. EzrinTD’s specific conformation reduces intermolecular interactions, allows binding to actin filaments, which reduces membrane tethering, and promotes ezrin binding to positively-curved membranes. While neither ezrinTD nor ezrinWT senses negative curvature alone, we demonstrate that interacting with curvature-sensing I-BAR-domain proteins facilitates ezrin enrichment in negatively-curved membrane protrusions. Overall, our work demonstrates that ezrin can tether membranes, or be targeted to curved membranes, depending on conformations and interactions with actin and curvature-sensing binding partners.
AB - One challenge in cell biology is to decipher the biophysical mechanisms governing protein enrichment on curved membranes and the resulting membrane deformation. The ERM protein ezrin is abundant and associated with cellular membranes that are flat, positively or negatively curved. Using in vitro and cell biology approaches, we assess mechanisms of ezrin’s enrichment on curved membranes. We evidence that wild-type ezrin (ezrinWT) and its phosphomimetic mutant T567D (ezrinTD) do not deform membranes but self-assemble anti-parallelly, zipping adjacent membranes. EzrinTD’s specific conformation reduces intermolecular interactions, allows binding to actin filaments, which reduces membrane tethering, and promotes ezrin binding to positively-curved membranes. While neither ezrinTD nor ezrinWT senses negative curvature alone, we demonstrate that interacting with curvature-sensing I-BAR-domain proteins facilitates ezrin enrichment in negatively-curved membrane protrusions. Overall, our work demonstrates that ezrin can tether membranes, or be targeted to curved membranes, depending on conformations and interactions with actin and curvature-sensing binding partners.
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U2 - 10.7554/eLife.37262
DO - 10.7554/eLife.37262
M3 - Article
C2 - 30234483
AN - SCOPUS:85054416261
SN - 2050-084X
VL - 7
JO - eLife
JF - eLife
M1 - e37262
ER -