Actin and Dynamin2 Dynamics and Interplay During Clathrin-mediated Endocytosis

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2014 Jun 4

PMID 24891602

Citations 112

Authors

Alexandre Grassart

Aaron T Cheng

Sun Hae Hong

Fan Zhang

Nathan Zenzer

Yongmei Feng

David M Briner

Gregory D Davis

Dmitry Malkov

David G Drubin

Affiliations

Soon will be listed here.

Abstract

Clathrin-mediated endocytosis (CME) involves the recruitment of numerous proteins to sites on the plasma membrane with prescribed timing to mediate specific stages of the process. However, how choreographed recruitment and function of specific proteins during CME is achieved remains unclear. Using genome editing to express fluorescent fusion proteins at native levels and live-cell imaging with single-molecule sensitivity, we explored dynamin2 stoichiometry, dynamics, and functional interdependency with actin. Our quantitative analyses revealed heterogeneity in the timing of the early phase of CME, with transient recruitment of 2-4 molecules of dynamin2. In contrast, considerable regularity characterized the final 20 s of CME, during which ∼26 molecules of dynamin2, sufficient to make one ring around the vesicle neck, were typically recruited. Actin assembly generally preceded dynamin2 recruitment during the late phases of CME, and promoted dynamin recruitment. Collectively, our results demonstrate precise temporal and quantitative regulation of the dynamin2 recruitment influenced by actin polymerization.

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