Switch-like Arp2/3 activation upon WASP and WIP recruitment to an apparent threshold level by multivalent linker proteins in vivo

• Main Authors: Yidi Sun, Nicole T Leong, Tommy Jiang, Astou Tangara, Xavier Darzacq, David G Drubin
• Format: Article
• Language: English
• Published: eLife Sciences Publications Ltd 2017-08-01
• Series: eLife
• Subjects: WASP (yeast Las17); WIP (yeast Vrp1); Intersectin (Pan1-End3-Sla1); clathrin-mediated endocytosis; multivalent PRM-SH3 domain interactions; phase seperation
• Online Access: https://elifesciences.org/articles/29140

Actin-related protein 2/3 (Arp2/3) complex activation by nucleation promoting factors (NPFs) such as WASP, plays an important role in many actin-mediated cellular processes. In yeast, Arp2/3-mediated actin filament assembly drives endocytic membrane invagination and vesicle scission. Here we used genetics and quantitative live-cell imaging to probe the mechanisms that concentrate NPFs at endocytic sites, and to investigate how NPFs regulate actin assembly onset. Our results demonstrate that SH3 (Src homology 3) domain-PRM (proline-rich motif) interactions involving multivalent linker proteins play central roles in concentrating NPFs at endocytic sites. Quantitative imaging suggested that productive actin assembly initiation is tightly coupled to accumulation of threshold levels of WASP and WIP, but not to recruitment kinetics or release of autoinhibition. These studies provide evidence that WASP and WIP play central roles in establishment of a robust multivalent SH3 domain-PRM network in vivo, giving actin assembly onset at endocytic sites a switch-like behavior.