Clathrin-adaptor ratio and membrane tension regulate the flat-to-curved transition of the clathrin coat during endocytosis

Nat Commun. 2018 Mar 16;9(1):1109. doi: 10.1038/s41467-018-03533-0.

Abstract

Although essential for many cellular processes, the sequence of structural and molecular events during clathrin-mediated endocytosis remains elusive. While it was long believed that clathrin-coated pits grow with a constant curvature, it was recently suggested that clathrin first assembles to form flat structures that then bend while maintaining a constant surface area. Here, we combine correlative electron and light microscopy and mathematical growth laws to study the ultrastructural rearrangements of the clathrin coat during endocytosis in BSC-1 mammalian cells. We confirm that clathrin coats initially grow flat and demonstrate that curvature begins when around 70% of the final clathrin content is acquired. We find that this transition is marked by a change in the clathrin to clathrin-adaptor protein AP2 ratio and that membrane tension suppresses this transition. Our results support the notion that BSC-1 mammalian cells dynamically regulate the flat-to-curved transition in clathrin-mediated endocytosis by both biochemical and mechanical factors.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • Clathrin / metabolism*
  • Coated Pits, Cell-Membrane / metabolism
  • Coated Pits, Cell-Membrane / ultrastructure*
  • Endocytosis / physiology*
  • Fatty Acid-Binding Proteins / metabolism*
  • Microscopy, Electron, Transmission
  • Osmotic Pressure / physiology*

Substances

  • Clathrin
  • Fatty Acid-Binding Proteins