Cells simultaneously assemble functionally diverse actin cytoskeleton filaments with distinct architectures and dynamics to drive fundamental processes such as polarization, endocytosis, motility and division. Multiple actin filament-based structures must self-organize within a single crowded cytoplasm to maintain organization and functional specificity. Numerous actin-binding proteins with different properties regulate actin assembly and organization. Cells remain poised to assemble actin filaments at the right time and place in response to signals because they maintain a large pool of actin monomers bound to the small actin-binding protein profilin. Profilin prevents unwanted spontaneous actin assembly by inhibiting nucleation, so factors are required to rapidly stimulate actin assembly at the right time and place in response to signals. Most new actin filaments are produced by an expanding list of nucleation and elongation factors that establish diverse actin filament architectures required for different processes. Formin utilizes a conceptually novel mechanism to stimulate the assembly of rapidly elongating long-straight filaments that are often pulled by myosin motors to generate contractile forces, or to provide polarized tracks and/or scaffolds that establish/maintain cell polarity and filopodia extension. Ena/VASP also promotes the elongation of long-straight filaments for extending filopodia at the leading edge of migrating cells. Conversely, Arp2/3 complex initiates the assembly of short-branched filaments that provide pushing forces. A major unresolved question is to determine how diverse nucleation factors work alone and in combination to assemble profilin-actin into functionally diverse actin filaments within a crowded common cytoplasm. Our hypothesis is that actin monomers are limiting, and therefore competition for profilin-actin by nucleation factors plays a critical role in maintaining a balance between different actin filament-based structures within a crowded cytoplasm. Profilin not only facilitates formin- and Ena/VASP-mediated actin filament elongation, but also allows formin to successfully compete with an excess of Arp2/3 complex for actin monomers. Our research goals are to determine the essential cellular role(s) of profilin, and follow up on progress made in our previous grant to investigate the precise mechanisms by which formins utilize profilin-actin for rapid actin filament elongation (Aim I). We are also expanding into the high impact area of investigating how formin competes with Arp2/3 complex for profilin-actin to drive different processes (Aim II), and how formin competes and/or cooperates with Ena/VASP to assemble actin filaments for filopodia (Aim III).
Cells require carefully controlled regulation of the actin filament cytoskeleton to drive fundamental processes such as division, polarization, motility, and thus, survival. We are studying how cells utilize diverse sets of complementary proteins to simultaneously organize the actin cytoskeleton for different processes. Understanding how these proteins collectively operate is critical to understanding how both healthy and diseased cells function, which will increase our ability to prevent and/or treat conditions such as birth defects and cancer.
|Winkelman, Jonathan D; Suarez, Cristian; Hocky, Glen M et al. (2016) Fascin- and Î±-Actinin-Bundled Networks Contain Intrinsic Structural Features that Drive Protein Sorting. Curr Biol 26:2697-2706|
|Zimmermann, Dennis; Morganthaler, Alisha N; Kovar, David R et al. (2016) In Vitro Biochemical Characterization of Cytokinesis Actin-Binding Proteins. Methods Mol Biol 1369:151-79|
|Suarez, Cristian; Kovar, David R (2016) Internetwork competition for monomers governs actin cytoskeleton organization. Nat Rev Mol Cell Biol 17:799-810|
|Bezanilla, Magdalena; Gladfelter, Amy S; Kovar, David R et al. (2015) Cytoskeletal dynamics: a view from the membrane. J Cell Biol 209:329-37|
|Bestul, Andrew J; Christensen, Jenna R; Grzegorzewska, Agnieszka P et al. (2015) Fission yeast profilin is tailored to facilitate actin assembly by the cytokinesis formin Cdc12. Mol Biol Cell 26:283-93|
|Suarez, Cristian; Carroll, Robert T; Burke, Thomas A et al. (2015) Profilin regulates F-actin network homeostasis by favoring formin over Arp2/3 complex. Dev Cell 32:43-53|
|Rotty, Jeremy D; Wu, Congying; Haynes, Elizabeth M et al. (2015) Profilin-1 serves as a gatekeeper for actin assembly by Arp2/3-dependent and -independent pathways. Dev Cell 32:54-67|
|Heisler, David B; Kudryashova, Elena; Grinevich, Dmitry O et al. (2015) ACTIN-DIRECTED TOXIN. ACD toxin-produced actin oligomers poison formin-controlled actin polymerization. Science 349:535-9|
|Zimmermann, Dennis; Santos, Alicja; Kovar, David R et al. (2015) Actin age orchestrates myosin-5 and myosin-6 run lengths. Curr Biol 25:2057-62|
|Burke, Thomas A; Christensen, Jenna R; Barone, Elisabeth et al. (2014) Homeostatic actin cytoskeleton networks are regulated by assembly factor competition for monomers. Curr Biol 24:579-85|
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