title: Diacylglycerol Guides the Hopping of Clathrin-Coated Pits along Microtubules for Exo-Endocytosis Coupling.
abstract: Many receptor-mediated endocytic processes are mediated by constitutive budding of clathrin-coated pits (CCPs) at spatially randomized sites before slowly pinching off from the plasma membrane (60-100 s). In contrast, clathrin-mediated endocytosis (CME) coupled with regulated exocytosis in excitable cells occurs at peri-exocytic sites shortly after vesicle fusion (∼10 s). The molecular mechanism underlying this spatiotemporal coupling remains elusive. We show that coupled endocytosis makes use of pre-formed CCPs, which hop to nascent fusion sites nearby following vesicle exocytosis. A dynamic cortical microtubular network, anchored at the cell surface by the cytoplasmic linker-associated protein on microtubules and the LL5β/ELKS complex on the plasma membrane, provides the track for CCP hopping. Local diacylglycerol gradients generated upon exocytosis guide the direction of hopping. Overall, the CCP-cytoskeleton-lipid interaction demonstrated here mediates exocytosis-coupled fast recycling of both plasma membrane and vesicular proteins, and it is required for the sustained exocytosis during repetitive stimulations.
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Tianyi
authors.initials: T
authors.lastname: Yuan
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Liu
authors.affiliation: National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Yongdeng
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Lisi
authors.initials: L
authors.lastname: Wei
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Shiqun
authors.initials: S
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Xiaolu
authors.initials: X
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Xiaoshuai
authors.initials: X
authors.lastname: Huang
authors.affiliation: UMR144 CNRS/UPMC, Institut Curie, 75005 Paris, France.
authors.forename: Jerome
authors.initials: J
authors.lastname: Boulanger
authors.affiliation: PICT Cell and Tissue Imaging Core Facility, Institut Curie, 75005 Paris, France; Roper Scientific SAS, 91017 Evry, France.
authors.forename: Charles
authors.initials: C
authors.lastname: Gueudry
authors.affiliation: National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Jingze
authors.initials: J
authors.lastname: Lu
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Lihan
authors.initials: L
authors.lastname: Xie
authors.affiliation: National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Du
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Weijian
authors.initials: W
authors.lastname: Zong
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Yang
authors.affiliation: UMR144 CNRS/UPMC, Institut Curie, 75005 Paris, France; PICT Cell and Tissue Imaging Core Facility, Institut Curie, 75005 Paris, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Salamero
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China. Electronic address: yanmeiliu@pku.edu.cn.
authors.forename: Yanmei
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing 100871, China; National Center for Nanoscience and Technology, Beijing 100871, China. Electronic address: lychen@pku.edu.cn.
authors.forename: Liangyi
authors.initials: L
authors.lastname: Chen
mesh_terms: D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000502:physiology*; D003599:Cytoskeleton / Q000378:metabolism; D004075:Diglycerides / Q000378:metabolism*; D004594:Electrophysiology; D005089:Exocytosis / Q000502:physiology*; D007091:Image Processing, Computer-Assisted; D007340:Insulinoma / Q000378:metabolism* / Q000473:pathology; D008561:Membrane Fusion / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism; D008870:Microtubules / Q000502:physiology*; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D014407:Tumor Cells, Cultured
chemical_list: C011439:1,2-diacylglycerol; D002966:Clathrin; D004075:Diglycerides; D008565:Membrane Proteins