TY - JOUR
T1 - Dynamin is primed at endocytic sites for ultrafast endocytosis
AU - Imoto, Yuuta
AU - Raychaudhuri, Sumana
AU - Ma, Ye
AU - Fenske, Pascal
AU - Sandoval, Eduardo
AU - Itoh, Kie
AU - Blumrich, Eva Maria
AU - Matsubayashi, Hideaki T.
AU - Mamer, Lauren
AU - Zarebidaki, Fereshteh
AU - Söhl-Kielczynski, Berit
AU - Trimbuch, Thorsten
AU - Nayak, Shraddha
AU - Iwasa, Janet H.
AU - Liu, Jian
AU - Wu, Bin
AU - Ha, Taekjip
AU - Inoue, Takanari
AU - Jorgensen, Erik M.
AU - Cousin, Michael A.
AU - Rosenmund, Christian
AU - Watanabe, Shigeki
N1 - Funding Information:
We thank Pietro De Camilli and Shawn Ferguson for sharing reagents and mice; Ira Milosevic for advice on antibodies; and Geraldine Seydoux, Philip Robinson and Jie Xiao for discussion. We are also indebted to M. Delanoy; B. Smith and Hoku West-Foyle at the Johns Hopkins Microscopy Facility and Sebastian Markert for technical assistance in electron and optical microscopy; Tyler Ogunmowo; Quan Gan and Sydney Brown for animal husbandry and the preparation of cells; Yuta Nihongaki for the helpful discussion on in vitro protein assay; Grant F. Kusick for editing of the manuscript; and Kyu Young Han for the initial building of the STED microscope. S.W. and this work were supported by start-up funds from the Johns Hopkins University School of Medicine, Johns Hopkins Discovery funds, Johns Hopkins Catalyst award, the National Science Foundation (1727260), and the National Institutes of Health (1DP2 NS111133-01 and 1R01 NS105810-01A1) awarded to S.W. and German research council-funded grants CRG958/A5, Exc257, and the Reinhard Koselleck project awarded to C.R. S.W. is an Alfred P. Sloan fellow, a McKnight Foundation Scholar, and a Klingenstein and Simons Foundation scholar. Y.I. was supported by JSPS. M.A.C. is supported by the Wellcome Trust (204954/Z/16/Z). E.M.J. is supported by the NIH grant NS034307 and is an investigator of the Howard Hughes Medical Institute. T.I. and J.L. are supported by National Science Foundation (NSF 2148534). Y.I. S.R. Y.M. and S.W. conceived the study and designed the experiments. C.R. and E.M.J. oversaw the pilot phase of the project. S.W. oversaw the overall research. Y.I. S.R. P.F. L.M. F.Z. and B.S.-K. performed the freezing experiments. Y.I. S.R. Y.M. E.S. K.I. and S.W. collected and analyzed the data. Y.M. set up the STED microscope and wrote the analysis codes for STED and lattice-light sheet microscopy (LLSM) images. Y.M. and B.W. built LLSM. S.N. and J.H.I. generated the scientific animation. E.-M.B. and M.A.C. performed the pull-down assay. Y.I. and H.T.M. prepared proteins for in vitro experiments. Y.I. S.R. P.F. and K.I. prepared neuron cultures. Y.I. S.R. E.S. K.I. H.T.M. and T.T. generated DNA constructs for the study. J.L. carried out the theoretical estimation. Y.I. and S.W. wrote the manuscript. All authors contributed to editing of the manuscript. B.W. T.H. T.I. C.R. and S.W. funded the research. The authors declare no competing interests.
Funding Information:
We thank Pietro De Camilli and Shawn Ferguson for sharing reagents and mice; Ira Milosevic for advice on antibodies; and Geraldine Seydoux, Philip Robinson and Jie Xiao for discussion. We are also indebted to M. Delanoy; B. Smith and Hoku West-Foyle at the Johns Hopkins Microscopy Facility and Sebastian Markert for technical assistance in electron and optical microscopy; Tyler Ogunmowo; Quan Gan and Sydney Brown for animal husbandry and the preparation of cells; Yuta Nihongaki for the helpful discussion on in vitro protein assay; Grant F. Kusick for editing of the manuscript; and Kyu Young Han for the initial building of the STED microscope. S.W. and this work were supported by start-up funds from the Johns Hopkins University School of Medicine , Johns Hopkins Discovery funds , Johns Hopkins Catalyst award , the National Science Foundation ( 1727260 ), and the National Institutes of Health ( 1DP2 NS111133-01 and 1R01 NS105810-01A1 ) awarded to S.W. and German research council -funded grants CRG958/A5 , Exc257 , and the Reinhard Koselleck project awarded to C.R. S.W. is an Alfred P. Sloan fellow, a McKnight Foundation Scholar , and a Klingenstein and Simons Foundation scholar. Y.I. was supported by JSPS . M.A.C. is supported by the Wellcome Trust ( 204954/Z/16/Z ). E.M.J. is supported by the NIH grant NS034307 and is an investigator of the Howard Hughes Medical Institute. T.I. and J.L. are supported by National Science Foundation (NSF 2148534 ).
Publisher Copyright:
© 2022 The Author(s)
PY - 2022/9/7
Y1 - 2022/9/7
N2 - Dynamin mediates fission of vesicles from the plasma membrane during endocytosis. Typically, dynamin is recruited from the cytosol to endocytic sites, requiring seconds to tens of seconds. However, ultrafast endocytosis in neurons internalizes vesicles as quickly as 50 ms during synaptic vesicle recycling. Here, we demonstrate that Dynamin 1 is pre-recruited to endocytic sites for ultrafast endocytosis. Specifically, Dynamin 1xA, a splice variant of Dynamin 1, interacts with Syndapin 1 to form molecular condensates on the plasma membrane. Single-particle tracking of Dynamin 1xA molecules confirms the liquid-like property of condensates in vivo. When Dynamin 1xA is mutated to disrupt its interaction with Syndapin 1, the condensates do not form, and consequently, ultrafast endocytosis slows down by 100-fold. Mechanistically, Syndapin 1 acts as an adaptor by binding the plasma membrane and stores Dynamin 1xA at endocytic sites. This cache bypasses the recruitment step and accelerates endocytosis at synapses.
AB - Dynamin mediates fission of vesicles from the plasma membrane during endocytosis. Typically, dynamin is recruited from the cytosol to endocytic sites, requiring seconds to tens of seconds. However, ultrafast endocytosis in neurons internalizes vesicles as quickly as 50 ms during synaptic vesicle recycling. Here, we demonstrate that Dynamin 1 is pre-recruited to endocytic sites for ultrafast endocytosis. Specifically, Dynamin 1xA, a splice variant of Dynamin 1, interacts with Syndapin 1 to form molecular condensates on the plasma membrane. Single-particle tracking of Dynamin 1xA molecules confirms the liquid-like property of condensates in vivo. When Dynamin 1xA is mutated to disrupt its interaction with Syndapin 1, the condensates do not form, and consequently, ultrafast endocytosis slows down by 100-fold. Mechanistically, Syndapin 1 acts as an adaptor by binding the plasma membrane and stores Dynamin 1xA at endocytic sites. This cache bypasses the recruitment step and accelerates endocytosis at synapses.
KW - Dyn1xA
KW - Dynamin
KW - Dynamin splice variants
KW - Syndapin
KW - endocytosis
KW - flash-and-freeze
KW - liquid condensates
KW - phase separation
KW - synaptic vesicle recycling
KW - ultrafast endocytosis
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U2 - 10.1016/j.neuron.2022.06.010
DO - 10.1016/j.neuron.2022.06.010
M3 - Article
C2 - 35809574
AN - SCOPUS:85135296462
SN - 0896-6273
VL - 110
SP - 2815-2835.e13
JO - Neuron
JF - Neuron
IS - 17
ER -