A Pan1/End3/Sla1 complex links Arp2/3-mediated actin assembly to sites of clathrin-mediated endocytosis

Molecular Biology of the Cell
Yidi SunDavid G Drubin

Abstract

More than 60 highly conserved proteins appear sequentially at sites of clathrin-mediated endocytosis in yeast and mammals. The yeast Eps15-related proteins Pan1 and End3 and the CIN85-related protein Sla1 are known to interact with each other in vitro, and they all appear after endocytic-site initiation but before endocytic actin assembly, which facilitates membrane invagination/scission. Here we used live-cell imaging in parallel with genetics and biochemistry to explore comprehensively the dynamic interactions and functions of Pan1, End3, and Sla1. Our results indicate that Pan1 and End3 associate in a stable manner and appear at endocytic sites before Sla1. The End3 C-terminus is necessary and sufficient for its cortical localization via interaction with Pan1, whereas the End3 N-terminus plays a crucial role in Sla1 recruitment. We systematically examined the dynamic behaviors of endocytic proteins in cells in which Pan1 and End3 were simultaneously eliminated, using the auxin-inducible degron system. The results lead us to propose that endocytic-site initiation and actin assembly are separable processes linked by a Pan1/End3/Sla1 complex. Finally, our study provides mechanistic insights into how Pan1 and End3 function with ...Continue Reading

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Citations

Nov 12, 2015·Developmental Cell·Eric B LewellynDavid G Drubin
Apr 17, 2016·Journal of Cell Science·Rebecca LuYidi Sun
Feb 8, 2018·Nature Reviews. Molecular Cell Biology·Marko Kaksonen, Aurélien Roux
Apr 21, 2018·Science·Elena KuzminChad L Myers
Feb 24, 2018·Frontiers in Cellular and Infection Microbiology·Hagit Bar-YosefDaniel Kornitzer
Mar 16, 2018·G3 : Genes - Genomes - Genetics·Kristie WrasmanBeverly Wendland
Feb 28, 2020·Journal of Fungi·Christiane RollenhagenSamuel A Lee
Jan 20, 2019·The Journal of Cell Biology·Ross T A Pedersen, David G Drubin
Nov 19, 2018·Biophysics Reviews·Kenneth S Gerien, Jian-Qiu Wu
Jun 18, 2020·Cells·Pika Miklavc, Manfred Frick
Sep 11, 2020·Journal of Cell Science·Thomas O TolsmaSantiago M Di Pietro
Oct 15, 2020·The Journal of Cell Biology·Ross T A PedersenDavid G Drubin

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Methods Mentioned

BETA
gel filtration
Immunoprecipitation
fluorescence
cross-correlation spectroscopy

Software Mentioned

Imaris
Icy image
ImageJ
Icy

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