Small GTPases and BAR Domain Proteins Regulate Branched Actin Polymerisation for Clathrin and Dynamin-independent Endocytosis

Overview

Journal Nat Commun

Specialty Biology

Date 2018 May 11

PMID 29743604

Citations 45

Authors

Mugdha Sathe

Gayatri Muthukrishnan

James Rae

Andrea Disanza

Mukund Thattai

Giorgio Scita

Robert G Parton

Satyajit Mayor

Affiliations

Soon will be listed here.

Abstract

Using real-time TIRF microscopy imaging, we identify sites of clathrin and dynamin-independent CLIC/GEEC (CG) endocytic vesicle formation. This allows spatio-temporal localisation of known molecules affecting CG endocytosis; GBF1 (a GEF for ARF1), ARF1 and CDC42 which appear sequentially over 60 s, preceding scission. In an RNAi screen for BAR domain proteins affecting CG endocytosis, IRSp53 and PICK1, known interactors of CDC42 and ARF1, respectively, were selected. Removal of IRSp53, a negative curvature sensing protein, abolishes CG endocytosis. Furthermore, the identification of ARP2/3 complex at CG endocytic sites, maintained in an inactive state reveals a function for PICK1, an ARP2/3 inhibitor. The spatio-temporal sequence of the arrival and disappearance of the molecules suggest a mechanism for a clathrin and dynamin-independent endocytic process. Coincident with the loss of PICK1 by GBF1-activated ARF1, CDC42 recruitment leads to the activation of IRSp53 and the ARP2/3 complex, resulting in a burst of F-actin polymerisation potentially powering scission.

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