Dynamic instability of clathrin assembly provides proofreading control for endocytosis

Clathrin-mediated endocytosis depends on the formation of functional clathrin-coated pits that recruit cargos and mediate the uptake of those cargos into the cell. However, it remains unclear whether the cargos in the growing clathrin-coated pits are actively monitored by the coat assembly machinery. Using a cell-free reconstitution system, we report that clathrin coat formation and cargo sorting can be uncoupled, indicating that a checkpoint is required for functional cargo incorporation. We demonstrate that the ATPase Hsc70 and a dynamic exchange of clathrin during assembly are required for this checkpoint. In the absence of Hsc70 function, clathrin assembles into pits but fails to enrich cargo. Using single-molecule imaging, we further show that uncoating takes place throughout the lifetime of the growing clathrin-coated pits. Our results suggest that the dynamic exchange of clathrin, at the cost of the reduced overall assembly rates, primarily serves as a proofreading mechanism for quality control of endocytosis.

Source: Journal of Cell Biology - October 6, 2019 Category: Cytology Authors: Chen, Y., Yong, J., Martinez-Sanchez, A., Yang, Y., Wu, Y., De Camilli, P., Fernandez-Busnadiego, R., Wu, M. Tags: Membrane and Lipid Biology, Trafficking, Biochemistry, Biophysics Reports Source Type: research