Three-color single molecule imaging shows WASP detachment from Arp2/3 complex triggers actin filament branch formation.

Abstract:

:During cell locomotion and endocytosis, membrane-tethered WASP proteins stimulate actin filament nucleation by the Arp2/3 complex. This process generates highly branched arrays of filaments that grow toward the membrane to which they are tethered, a conflict that seemingly would restrict filament growth. Using three-color single-molecule imaging in vitro we revealed how the dynamic associations of Arp2/3 complex with mother filament and WASP are temporally coordinated with initiation of daughter filament growth. We found that WASP proteins dissociated from filament-bound Arp2/3 complex prior to new filament growth. Further, mutations that accelerated release of WASP from filament-bound Arp2/3 complex proportionally accelerated branch formation. These data suggest that while WASP promotes formation of pre-nucleation complexes, filament growth cannot occur until it is triggered by WASP release. This provides a mechanism by which membrane-bound WASP proteins can stimulate network growth without restraining it. DOI:http://dx.doi.org/10.7554/eLife.01008.001.

journal_name

Elife

journal_title

eLife

authors

Smith BA,Padrick SB,Doolittle LK,Daugherty-Clarke K,Corrêa IR Jr,Xu MQ,Goode BL,Rosen MK,Gelles J

doi

10.7554/eLife.01008

subject

Has Abstract

pub_date

2013-09-03 00:00:00

pages

e01008

issn

2050-084X

pii

01008

journal_volume

2

pub_type

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