In this Program Project Grant our overarching hypothesis is that alterations in the assembly states and mechanical properties of cytoskeletal IF, specifically the type III IF composed of vimentin (VIF), play important roles in regulating the micromechanical properties of cells in response to mechano- and chemo- signaling. These studies are critically important as IF are major elements of the cytoskeletal system of mammalian cells and yet their specific functions in cell motility remain unknown. The 6 Project Leaders, and their research aims are as follows: R. Goldman, Northwestern University, will determine how the vimentin IF (VIF) system changes in response to mechanical and chemical signals, and will purify and characterize the vimentin precursors that form as VIF assemble and disassemble. V. Gelfand, Northwestern University will determine the relationship between VIF with microtubules and actin filaments, identify the molecular motors responsible for translocating VIF precursors, and investigate how VIF affect cytoskeletal dynamics and microtubule (MT) dynamics. G. Danuser, Harvard University, will test the hypotheses that VIF network formation involves a spatially distributed assembly line, that this process involves molecular motors, and that VIF network assembly modulates assembly of MT and microfilaments (MF). D. Weitz, Harvard University, will determine the micromechanical properties of in vitro assembled VIF networks prepared from purified vimentin, native VIF networks isolated from cells and in living cells. P. Janmey, University of Pennsylvania, will determine how the VIF network contributes to the micromechanical properties of living cells and how this changes in response to substrate stiffness and external forces. P. Burkhard, University of Connecticut, will use biophysical techniques and X-ray crystallography to determine the structure and biophysical properties of the vimentin dimer and study the formation of VIF assembly intermediates. In all of these studies, the regulatory function of different posttranslational phosphorylation events will be examined.
Intermediate Filaments (IF) are structural proteins involved in determining cell shape, movement and mechanical integrity. However, their specific functions remain unknown. We propose studies aimed at understanding their specific functions in the movement and mechanical properties of cells. These basic studies will provide insights into their normal functions and will help explain their defective functions in the many diseases attributed to mutations in the genes encoding the different IF proteins.
|Charrier, Elisabeth E; Janmey, Paul A (2016) Mechanical Properties of Intermediate Filament Proteins. Methods Enzymol 568:35-57|
|Lin, Ni-Hsuan; Huang, Yu-Shan; Opal, Puneet et al. (2016) The role of gigaxonin in the degradation of the glial-specific intermediate filament protein GFAP. Mol Biol Cell 27:3980-3990|
|Lowery, Jason; Jain, Nikhil; Kuczmarski, Edward R et al. (2016) Abnormal intermediate filament organization alters mitochondrial motility in giant axonal neuropathy fibroblasts. Mol Biol Cell 27:608-16|
|Ridge, Karen M; Shumaker, Dale; Robert, AmÃ©lie et al. (2016) Methods for Determining the Cellular Functions of Vimentin Intermediate Filaments. Methods Enzymol 568:389-426|
|Gan, Zhuo; Ding, Liya; Burckhardt, Christoph J et al. (2016) Vimentin Intermediate Filaments Template Microtubule Networks to Enhance Persistence in Cell Polarity and Directed Migration. Cell Syst 3:252-263.e8|
|Robert, AmÃ©lie; Hookway, Caroline; Gelfand, Vladimir I (2016) Intermediate filament dynamics: What we can see now and why it matters. Bioessays 38:232-43|
|Utech, Stefanie; Prodanovic, Radivoje; Mao, Angelo S et al. (2015) Microfluidic Generation of Monodisperse, Structurally Homogeneous Alginate Microgels for Cell Encapsulation and 3D Cell Culture. Adv Healthc Mater 4:1628-33|
|Klein, Allon M; Mazutis, Linas; Akartuna, Ilke et al. (2015) Droplet barcoding for single-cell transcriptomics applied to embryonic stem cells. Cell 161:1187-201|
|KÃ¶ster, Sarah; Weitz, David A; Goldman, Robert D et al. (2015) Intermediate filament mechanics in vitro and in the cell: from coiled coils to filaments, fibers and networks. Curr Opin Cell Biol 32:82-91|
|Galie, P A; van Oosten, A; Chen, C S et al. (2015) Application of multiple levels of fluid shear stress to endothelial cells plated on polyacrylamide gels. Lab Chip 15:1205-12|
Showing the most recent 10 out of 38 publications