title: G-actin regulates rapid induction of actin nucleation by mDia1 to restore cellular actin polymers.
abstract: mDia1 belongs to the formin family of proteins that share FH1 and FH2 domains. Although formins play a critical role in the formation of many actin-based cellular structures, the physiological regulation of formin-mediated actin assembly within the cell is still unknown. Here we show that cells possess an acute actin polymer restoration mechanism involving mDia1. By using single-molecule live-cell imaging, we found that several treatments including low-dose G-actin-sequestering drugs and unpolymerizable actin mutants activate mDia1 to initiate fast directional movement. The FH2 region, the core domain for actin nucleation, is sufficient to respond to latrunculin B (LatB) to increase its actin nucleation frequency. Simulation analysis revealed an unexpected paradoxical effect of LatB that leads to a several fold increase in free G-actin along with an increase in total G-actin. These results indicate that in cells, the actin nucleation frequency of mDia1 is enhanced not only by Rho, but also strongly through increased catalytic efficiency of the FH2 domain. Consistently, frequent actin nucleation by mDia1 was found around sites of vigorous actin disassembly. Another major actin nucleator, the Arp2/3 complex, was not affected by the G-actin increase induced by LatB. Taken together, we propose that transient accumulation of G-actin works as a cue to promote mDia1-catalyzed actin nucleation to execute rapid reassembly of actin filaments.
authors.affiliation: Department of Pharmacology, Kyoto University Faculty of Medicine, Yoshida Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Chiharu
authors.initials: C
authors.lastname: Higashida
authors.forename: Shiro
authors.initials: S
authors.lastname: Suetsugu
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tsuji
authors.forename: James
authors.initials: J
authors.lastname: Monypenny
authors.forename: Shuh
authors.initials: S
authors.lastname: Narumiya
authors.forename: Naoki
authors.initials: N
authors.lastname: Watanabe
mesh_terms: D008841:Actin Cytoskeleton / Q000235:genetics / Q000378:metabolism*; D051376:Actin-Related Protein 2-3 Complex / Q000235:genetics / Q000378:metabolism*; D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D000080865:Formins; D051379:Mice; D017434:Protein Structure, Tertiary / Q000502:physiology; D053778:Thiazolidines / Q000494:pharmacology
chemical_list: D051376:Actin-Related Protein 2-3 Complex; D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002352:Carrier Proteins; C491464:Diap1 protein, mouse; D000080865:Formins; D053778:Thiazolidines; C037068:latrunculin B