article published yes Alexander J Johnson author 46A62C3A-F248-11E8-B48F-1D18A9856A870000-0002-2739-8843 Dana A Dahhan author Nataliia Gnyliukh author 390C1120-F248-11E8-B48F-1D18A9856A870000-0002-2198-0509 Walter Kaufmann author 3F99E422-F248-11E8-B48F-1D18A9856A870000-0001-9735-5315 Vanessa Zheden author 39C5A68A-F248-11E8-B48F-1D18A9856A870000-0002-9438-4783 Tommaso Costanzo author D93824F4-D9BA-11E9-BB12-F207E66974250000-0001-9732-3815 Pierre Mahou author Mónika Hrtyan author 45A71A74-F248-11E8-B48F-1D18A9856A87 Jie Wang author Juan L Aguilera Servin author 2A67C376-F248-11E8-B48F-1D18A9856A870000-0002-2862-8372 Daniël van Damme author Emmanuel Beaurepaire author Martin Loose author 462D4284-F248-11E8-B48F-1D18A9856A870000-0001-7309-9724 Sebastian Y Bednarek author Jiří Friml author 4159519E-F248-11E8-B48F-1D18A9856A870000-0002-8302-7596 JiFr department MaLo department EvBe department EM-Fac department NanoFab department Molecular mechanisms of endocytic cargo recognition in plants project Clathrin-mediated endocytosis is the major route of entry of cargos into cells and thus underpins many physiological processes. During endocytosis, an area of flat membrane is remodeled by proteins to create a spherical vesicle against intracellular forces. The protein machinery which mediates this membrane bending in plants is unknown. However, it is known that plant endocytosis is actin independent, thus indicating that plants utilize a unique mechanism to mediate membrane bending against high-turgor pressure compared to other model systems. Here, we investigate the TPLATE complex, a plant-specific endocytosis protein complex. It has been thought to function as a classical adaptor functioning underneath the clathrin coat. However, by using biochemical and advanced live microscopy approaches, we found that TPLATE is peripherally associated with clathrin-coated vesicles and localizes at the rim of endocytosis events. As this localization is more fitting to the protein machinery involved in membrane bending during endocytosis, we examined cells in which the TPLATE complex was disrupted and found that the clathrin structures present as flat patches. This suggests a requirement of the TPLATE complex for membrane bending during plant clathrin–mediated endocytosis. Next, we used in vitro biophysical assays to confirm that the TPLATE complex possesses protein domains with intrinsic membrane remodeling activity. These results redefine the role of the TPLATE complex and implicate it as a key component of the evolutionarily distinct plant endocytosis mechanism, which mediates endocytic membrane bending against the high-turgor pressure in plant cells. https://research-explorer.ista.ac.at/download/9887/10546/2021_PNAS_Johnson.pdf application/pdfno National Academy of Sciences2021 eng 1091-6490 34907016 00073641760004310.1073/pnas.2113046118 11851 https://research-explorer.ista.ac.at/record/14988 https://research-explorer.ista.ac.at/record/14510 https://doi.org/10.1101/2021.04.26.441441 Johnson, A. J., Dahhan, D. A., Gnyliukh, N., Kaufmann, W., Zheden, V., Costanzo, T., … Friml, J. (2021). The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis. <i>Proceedings of the National Academy of Sciences of the United States of America</i>. National Academy of Sciences. <a href="https://doi.org/10.1073/pnas.2113046118">https://doi.org/10.1073/pnas.2113046118</a> Johnson, Alexander J., et al. “The TPLATE Complex Mediates Membrane Bending during Plant Clathrin-Mediated Endocytosis.” <i>Proceedings of the National Academy of Sciences of the United States of America</i>, vol. 118, no. 51, e2113046118, National Academy of Sciences, 2021, doi:<a href="https://doi.org/10.1073/pnas.2113046118">10.1073/pnas.2113046118</a>. A. J. Johnson <i>et al.</i>, “The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis,” <i>Proceedings of the National Academy of Sciences of the United States of America</i>, vol. 118, no. 51. National Academy of Sciences, 2021. Johnson AJ, Dahhan DA, Gnyliukh N, Kaufmann W, Zheden V, Costanzo T, Mahou P, Hrtyan M, Wang J, Aguilera Servin JL, van Damme D, Beaurepaire E, Loose M, Bednarek SY, Friml J. 2021. The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis. Proceedings of the National Academy of Sciences of the United States of America. 118(51), e2113046118. Johnson AJ, Dahhan DA, Gnyliukh N, et al. The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis. <i>Proceedings of the National Academy of Sciences of the United States of America</i>. 2021;118(51). doi:<a href="https://doi.org/10.1073/pnas.2113046118">10.1073/pnas.2113046118</a> A.J. Johnson, D.A. Dahhan, N. Gnyliukh, W. Kaufmann, V. Zheden, T. Costanzo, P. Mahou, M. Hrtyan, J. Wang, J.L. Aguilera Servin, D. van Damme, E. Beaurepaire, M. Loose, S.Y. Bednarek, J. Friml, Proceedings of the National Academy of Sciences of the United States of America 118 (2021). Johnson, Alexander J, Dana A Dahhan, Nataliia Gnyliukh, Walter Kaufmann, Vanessa Zheden, Tommaso Costanzo, Pierre Mahou, et al. “The TPLATE Complex Mediates Membrane Bending during Plant Clathrin-Mediated Endocytosis.” <i>Proceedings of the National Academy of Sciences of the United States of America</i>. National Academy of Sciences, 2021. <a href="https://doi.org/10.1073/pnas.2113046118">https://doi.org/10.1073/pnas.2113046118</a>. 98872021-08-11T14:11:43Z2026-03-07T23:30:35Z