PT  - JOURNAL ARTICLE
AU  - Delia Bucher
AU  - Felix Frey
AU  - Kem A. Sochacki
AU  - Susann Kummer
AU  - Jan-Philip Bergeest
AU  - William J. Godinez
AU  - Hans-Georg Kräusslich
AU  - Karl Rohr
AU  - Justin W. Taraska
AU  - Ulrich S. Schwarz
AU  - Steeve Boulant
TI  - Flat-to-curved transition during clathrin-mediated endocytosis correlates with a change in clathrin-adaptor ratio and is regulated by membrane tension
AID  - 10.1101/162024
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 162024
4099  - http://biorxiv.org/content/early/2017/07/11/162024.short
4100  - http://biorxiv.org/content/early/2017/07/11/162024.full
AB  - Although essential for many cellular processes, the sequence of structural and molecular events during clathrin-mediated endocytosis remains elusive. While it was believed that clathrin-coated pits grow with a constant curvature, it was recently suggested that clathrin first assembles to form a flat structure and then bends while maintaining a constant surface area. Here, we combine correlative electron and light microscopy and mathematical modelling to quantify the sequence of ultrastructural rearrangements of the clathrin coat during endocytosis in mammalian cells. We confirm that clathrin-coated structures can initially grow flat and that lattice curvature does not show a direct correlation with clathrin coat assembly. We demonstrate that curvature begins when 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 model that mammalian cells dynamically regulate the flat-to-curved transition in clathrin-mediated endocytosis by both biochemical and mechanical factors.