In keeping with the tradition of this laboratory, the application consists of four relatively independent projects, connected here because they involve actin as opposed to myosin. The projects are as follows: (1) Characterize the effects of nucleotide and polyproline on actin-profilin interactions. Search for proteins that bind to the polyproline binding site of profilin. Use profilin mutants to test which biochemical interactions are important in vivo in fission yeast. (2) The Arp2/3 complex, identified as binding to profilin.
Aims are to sequence the other five proteins, to determine the structure of the complex and interactions between the subunits, and to reconstitute the complex from purified components. (3) To characterize the binding of actophorin (a cofilin/ADF relative) to actin, especially the severing of actin filaments by actophorin and to determine the effects in vivo of changing the actophorin concentration. (4) To test further a model in which alpha-actinin functions as a dynamic crosslinker of actin filaments in vitro.
| Espinoza-Sanchez, Sofia; Metskas, Lauren Ann; Chou, Steven Z et al. (2018) Conformational changes in Arp2/3 complex induced by ATP, WASp-VCA, and actin filaments. Proc Natl Acad Sci U S A 115:E8642-E8651 |
| Arasada, Rajesh; Sayyad, Wasim A; Berro, Julien et al. (2018) High-speed superresolution imaging of the proteins in fission yeast clathrin-mediated endocytic actin patches. Mol Biol Cell 29:295-303 |
| Aydin, Fikret; Courtemanche, Naomi; Pollard, Thomas D et al. (2018) Gating mechanisms during actin filament elongation by formins. Elife 7: |
| Friend, Janice E; Sayyad, Wasim A; Arasada, Rajesh et al. (2018) Fission yeast Myo2: Molecular organization and diffusion in the cytoplasm. Cytoskeleton (Hoboken) 75:164-173 |
| Fujiwara, Ikuko; Zweifel, Mark E; Courtemanche, Naomi et al. (2018) Latrunculin A Accelerates Actin Filament Depolymerization in Addition to Sequestering Actin Monomers. Curr Biol 28:3183-3192.e2 |
| Akamatsu, Matthew; Lin, Yu; Bewersdorf, Joerg et al. (2017) Analysis of interphase node proteins in fission yeast by quantitative and superresolution fluorescence microscopy. Mol Biol Cell 28:3203-3214 |
| Pollard, Thomas D (2017) Nine unanswered questions about cytokinesis. J Cell Biol 216:3007-3016 |
| Pollard, Thomas D (2017) What We Know and Do Not Know About Actin. Handb Exp Pharmacol 235:331-347 |
| Courtemanche, Naomi; Pollard, Thomas D; Chen, Qian (2016) Avoiding artefacts when counting polymerized actin in live cells with LifeAct fused to fluorescent proteins. Nat Cell Biol 18:676-83 |
| Pollard, Thomas D (2016) Actin and Actin-Binding Proteins. Cold Spring Harb Perspect Biol 8: |
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