Analysis of functional surfaces on the actin nucleation promoting factor Dip1 required for Arp2/3 complex activation and endocytic actin network assembly

Liu, Su-Ling; Narvaez-Ortiz, Heidy Y; Miner, Matt; Kiemel, Jack; Oberhelman, Nicholas J.; Watt, April C.; Wagner, Andrew R.; Luan, Qing; Helgeson, Luke A.; Nolen, Brad J.

doi:10.1016/j.jbc.2022.102019

Cited by 3 publications

(1 citation statement)

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“…Major actin nucleating proteins in cells include Arp2/3 complex and the formin family 43,44 . These nucleator proteins are recruited and regulated spatially and temporally by endogenous cellular proteins [45][46][47] and by proteins derived from pathogenic organisms 48,49 .…”

Section: Role Of Cp In Regulation Of Actin Polymerizationmentioning

confidence: 99%

Reconstitution of Arp2/3-Nucleated Actin Assembly with CP, V-1 and CARMIL

Mooren,

McConnell,

DeBrecht

et al. 2024

Preprint

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Actin polymerization is often associated with membrane proteins containing capping-protein-interacting (CPI) motifs, such as CARMIL, CD2AP, and WASHCAP/Fam21. CPI motifs bind directly to actin capping protein (CP), and this interaction weakens the binding of CP to barbed ends of actin filaments, lessening the ability of CP to functionally cap those ends. The protein V-1 / myotrophin binds to the F-actin binding site on CP and sterically blocks CP from binding barbed ends. CPI-motif proteins also weaken the binding between V-1 and CP, which decreases the inhibitory effects of V-1, thereby freeing CP to cap barbed ends. Here, we address the question of whether CPI-motif proteins on a surface analogous to a membrane lead to net activation or inhibition of actin assembly nucleated by Arp2/3 complex. Using reconstitution with purified components, we discovered that CARMIL at the surface promotes and enhances actin assembly, countering the inhibitory effects of V-1 and thus activating CP. The reconstitution involves the presence of an Arp2/3 activator on the surface, along with Arp2/3 complex, V-1, CP, profilin and actin monomers in solution, recreating key features of cell physiology.

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