title: Electrostatics control actin filament nucleation and elongation kinetics.
abstract: The actin cytoskeleton is a central mediator of cellular morphogenesis, and rapid actin reorganization drives essential processes such as cell migration and cell division. Whereas several actin-binding proteins are known to be regulated by changes in intracellular pH, detailed information regarding the effect of pH on the actin dynamics itself is still lacking. Here, we combine bulk assays, total internal reflection fluorescence microscopy, fluorescence fluctuation spectroscopy techniques, and theory to comprehensively characterize the effect of pH on actin polymerization. We show that both nucleation and elongation are strongly enhanced at acidic pH, with a maximum close to the pI of actin. Monomer association rates are similarly affected by pH at both ends, although dissociation rates are differentially affected. This indicates that electrostatics control the diffusional encounter but not the dissociation rate, which is critical for the establishment of actin filament asymmetry. A generic model of protein-protein interaction, including electrostatics, explains the observed pH sensitivity as a consequence of charge repulsion. The observed pH effect on actin in vitro agrees with measurements of Listeria propulsion in pH-controlled cells. pH regulation should therefore be considered as a modulator of actin dynamics in a cellular environment.
authors.affiliation: AG Cellular Dynamics and Cell Patterning, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany. alvaro.crevenna@cup.uni-muenchen.de
authors.forename: Alvaro H
authors.initials: AH
authors.lastname: Crevenna
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Naredi-Rainer
authors.forename: André
authors.initials: A
authors.lastname: Schönichen
authors.forename: Joachim
authors.initials: J
authors.lastname: Dzubiella
authors.forename: Diane L
authors.initials: DL
authors.lastname: Barber
authors.forename: Don C
authors.initials: DC
authors.lastname: Lamb
authors.forename: Roland
authors.initials: R
authors.lastname: Wedlich-Söldner
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D002645:Chickens; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008087:Listeria / Q000378:metabolism; D008956:Models, Chemical; D011817:Rabbits; D055672:Static Electricity