Cell motility, motor molecules, integrin-cytoskeleton interactions
The morphology of cells, organs and whole organisms is determined by the generation of forces on the immediate environment, which is either extracellular matrix or adjacent cells. We are currently engaged in studies to understand the detailed molecular mechanisms involved in a variety of phenomena from fertilization to brain function. Further, we are developing several new tools and protocols for measuring and applying forces at the molecular level, which are revealing many new aspects of how cells can both generate and respond to external forces.
Cell spreading, force generation and rigidity sensing
We have an effort underway to define quantitatively the steps involved in cell adhesion to and spreading on a matrix-coated surface. During spreading, the cell continuously tests the rigidity of the surface by applying forces to it via integrin adhesions at the cell edge. How cells develop forces, and how these forces are used to sense and transmit rigidity signals are both unknown. Our focus is on analyses of the steps in building the machinery used by fibroblasts to sense and transmit rigidity signals. We are using nanofabricated surfaces to track the recruitment of proteins to nascent adhesions and determine which proteins are essential for adhesion assembly and force production. We are also combining elastic micropillars together with high-speed microscopy to study the kinetics of force production on different rigidities. Using a variety of mouse fibroblast cell lines that are missing critical proteins will enable us to generate a working model of the process of cell spreading and force generation that is consistent with previous studies as well as our findings.
Mechanosensing in myofibrillogenesis
The aim of our research is to understand how cells assemble a force sensing machinery to probe the local microenvironment. The machinery is based on actomyosin arrays, which are connected to the substrate via surface receptors. We are investigating which proteins guide the assembly of the actin filaments that are formed from the surface receptors in the context of early cell spreading and during the initial steps of muscle differentiation. To answer these questions, we fabricate substrates with defined patterns and rigidities and analyze protein localization with highest spatial and temporal resolution.
Mechanotransduction at the immunological synapse
We have a collaborative effort underway to define quantitatively the steps involved in the formation of the immunological synapse between a T cell and an antigen presenting cell. Our aim is to understand how receptor-ligand interactions at the immunological synapse trigger adhesion, migration, and ultimately activation of naïve T cells. We are investigating the role of several candidate molecules as mechanosensors at the immunological synapse. Supported planar bilayers mimicking antigen presenting cells allow us to study early molecular events of immunological synapse formation with a high spatial and temporal resolution.
- Lynch CD, Lazar AM, Iskratsch T, Zhang X, Sheetz MP (2013) Endoplasmic spreading requires coalescence of vimentin intermediate filaments at force-bearing adhesions. Mol Biol Cell24(1): 21-30.
- Moore SW, Zhang X, Lynch CD, Sheetz MP. (2012) Netrin-1 attracts axons through FAK-dependent mechanotransduction. J Neurosci 32(34): 11574-85.
- Mathur A, Moore SW, Sheetz MP, Hone J. (2012) The role of feature curvature in contact guidance. Acta Biomater 8(7): 2595-601.
- Roca-Cusachs P, Iskratsch T, Sheetz MP. (2012) Finding the weakest link: exploring integrin-mediated mechanical molecular pathways J Cell Sci 125(Pt 13): 3025-38.
- Ghassemi, S., Meacci, G., Liu, S., Gondarenko, A. A., Mathur, A., Roca-Cusachs, P., Sheetz, M. P., Hone, J (2012) Cells test substrate rigidity by local contractions on submicrometer pillars. Proc Natl Acad Sci U S A 109(14): 5328-5333.
- Kumari S, Vardhana S, Cammer M, Curado S, Santos L, Sheetz MP, Dustin ML. (2012) T Lymphocyte Myosin IIA is Required for Maturation of the Immunological Synapse, Front. Immunol3: 230.
- Moore SW, Sheetz MP. (2011) Biophysics of substrate interaction: influence on neural motility, differentiation, and repair. Dev Neurobiol 71(11): 1090-101.
- Palma M, Abramson JJ, Gorodetsky AA, Penzo E, Gonzalez RL Jr, Sheetz MP, Nuckolls C, Hone J, Wind SJ. (2011) Selective biomolecular nanoarrays for parallel single-molecule investigations. J Am Chem Soc 133(20): 7656-9.
- Lynch CD, Gauthier NC, Biais N, Lazar AM, Roca-Cusachs P, Yu CH, Sheetz MP. (2011) Filamin depletion blocks endoplasmic spreading and destabilizes force-bearing adhesions. Mol Biol Cell 22(8): 1263-73.
- Palma M, Abramson JJ, Gorodetsky AA, Nuckolls C, Sheetz MP, Wind SJ, Hone J. (2011) Controlled confinement of DNA at the nanoscale: nanofabrication and surface bio-functionalization.Methods Mol Biol 749: 169-85.
- Moore SW, Roca-Cusachs P, Sheetz MP. (2010) Stretchy proteins on stretchy substrates: the important elements of integrin-mediated rigidity sensing. Dev Cell 19(2): 194-206.
- Biais N, Higashi DL, Brujic J, So M, Sheetz MP. (2010) Force-dependent polymorphism in type IV pili reveals hidden epitopes. Proc Natl Acad Sci U S A 107(25): 11358-63.
- Rossier OM, Gauthier N, Biais N, Vonnegut W, Fardin MA, Avigan P, Heller ER, Mathur A, Ghassemi S, Koeckert MS, Hone JC, Sheetz MP. (2010) Force generated by actomyosin contraction builds bridges between adhesive contacts. EMBO J29(6): 1055-68.
- Cai Y, Rossier O, Gauthier NC, Biais N, Fardin MA, Zhang X, Miller LW, Ladoux B, Cornish VW, Sheetz MP. (2010) Cytoskeletal coherence requires myosin-IIA contractility. J Cell Sci 123(Pt 3): 413-23.
- Moore SW, Biais N, Sheetz MP. (2009) Traction on immobilized netrin-1 is sufficient to reorient axons. Science 325(5937): 166.
- Ghassemi S, Rossier O, Sheetz MP, Wind SJ, Hone J. (2009) Gold-Tipped Elastomeric Pillars for Cellular Mechanotransduction. J Vac Sci Technol B Microelectron Nanometer Struct Process Meas Phenom 27(6): 3088-3091.
- del Rio, A., R. Perez-Jimenez, R. Liu, P. Roca-Cusachs, J.M. Fernandez, and M.P. Sheetz. (2009) Stretching single talin rod molecules activates vinculin binding. Science 323: 638-41.
- Biais, N., B. Ladoux, D. Higashi, M. So, and M. Sheetz. (2008) Cooperative retraction of bundled type IV pili enables nanonewton force generation. PLoS Biol 6: e87.
- Ghassemi S, Biais N, Maniura K, Wind SJ, Sheetz MP, Hone J. (2008) Fabrication of elastomer Pillar Arrays with Modulated Stiffness for Cellular Force Measurements. J Vac Sci Technol B Microelectron Nanometer Struct Process Meas Phenom. 26(6): 2549-2553.
- von Wichert G, Krndija D, Schmid H, von Wichert G, Haerter G, Adler G, Seufferlein T, Sheetz MP. (2008) Focal adhesion kinase mediates defects in the force-dependent reinforcement of initial integrin-cytoskeleton linkages in metastatic colon cancer cell lines. Eur J Cell Biol 87(1): 1-16.
- Perez TD, Tamada M, Sheetz MP, Nelson WJ (2008) Immediate-early signaling induced by E-cadherin engagement and adhesion J Biol Chem 283(8): 5014-22.
- Zhang X, Jiang G, Cai Y, Monkley SJ, Critchley DR, Sheetz MP. (2008) Talin depletion reveals independence of initial cell spreading from integrin activation and traction. Nat Cell Biol
- Kostic A, Sap J, Sheetz MP. (2007) RPTPalpha is required for rigidity-dependent inhibition of extension and differentiation of hippocampal neurons J Cell Sci 120(Pt 21): 3895-904.
- Sims TN, Soos TJ, Xenias HS, Dubin-Thaler B, Hofman JM, Waite JC, Cameron TO, Thomas VK, Varma R, Wiggins CH, Sheetz MP, Littman DR, Dustin ML (2007) Opposing effects of PKCtheta and WASp on symmetry breaking and relocation of the immunological synapse. Cell 129(4): 773-85.
- Boldogh IR, Pon LA, Sheetz MP, De Vos KJ. (2007) Cell-free assays for mitochondria-cytoskeleton interactions Methods Cell Biol 80: 683-706.
- Calloway NT, Choob M, Sanz A, Sheetz MP, Miller LW, Cornish VW (2007) Optimized fluorescent trimethoprim derivatives for in vivo protein labeling. Chembiochem 7;8(7): 767-74.
- De Vos KJ, Sheetz MP (2007) Visualization and quantification of mitochondrial dynamics in living animal cells. Methods Cell Biol 80: 627-82.
- Tran AD, Marmo TP, Salam AA, Che S, Finkelstein E, Kabarriti R, Xenias HS, Mazitschek R, Hubbert C, Kawaguchi Y, Sheetz MP, Yao TP, Bulinski JC. (2007) HDAC6 deacetylation of tubulin modulates dynamics of cellular adhesions. J Cell Sci J Cell Sci120( 8): 1469-79..
- Ahmed I, Ponery AS, Nur-E-Kamal A, Kamal J, Meshel AS, Sheetz MP, Schindler M, Meiners S. (2007) Morphology, cytoskeletal organization, and myosin dynamics of mouse embryonic fibroblasts cultured on nanofibrillar surfaces. Mol Cell Biochem 301(1-2): 241-9.
- Giannone G, Dubin-Thaler BJ, Rossier O, Cai Y, Chaga O, Jiang G, Beaver W, Döbereiner HG, Freund Y, Borisy G, Sheetz MP. (2007) Lamellipodial actin mechanically links myosin activity with adhesion-site formation. Cell 128(3): 561-75.
- Tamada M, Perez TD, Nelson WJ, Sheetz MP. (2007) Two distinct modes of myosin assembly and dynamics during epithelial wound closure J Cell Biol 176(1): 27-33.
- Sawada Y, Tamada M, Dubin-Thaler BJ, Cherniavskaya O, Sakai R, Tanaka S, Sheetz MP (2006) Force sensing by mechanical extension of the Src family kinase substrate p130Cas. Cell 127(5): 1015-26.
- Cai Y, Biais N, Giannone G, Tanase M, Jiang G, Hofman JM, Wiggins CH, Silberzan P, Buguin A, Ladoux B, Sheetz MP. (2006) Nonmuscle myosin IIA-dependent force inhibits cell spreading and drives F-actin flow. Biophys J 91(10): 3907-20.
- Döbereiner HG, Dubin-Thaler BJ, Hofman JM, Xenias HS, Sims TN, Giannone G, Dustin ML, Wiggins CH, Sheetz MP (2006) ustin ML, Wiggins CH, Sheetz MP. Lateral membrane waves constitute a universal dynamic pattern of motile cells. Phys Rev Lett 97(3): 038102.
- Sheetz MP, Sable JE, Döbereiner HG. (2006) Continuous membrane-cytoskeleton adhesion requires continuous accommodation to lipid and cytoskeleton dynamics. Annu Rev Biophys Biomol Struct 35: 417-34.
- Miller KE, Sheetz MP (2006) Direct evidence for coherent low velocity axonal transport of mitochondria. J Cell Biol 173(3): 373-81.
- Kostic A, Sheetz MP (2006) Fibronectin rigidity response through Fyn and p130Cas recruitment to the leading edge. Mol Biol Cell 17(6): 2684-95.
- Ada-Nguema AS, Xenias H, Hofman JM, Wiggins CH, Sheetz MP, Keely PJ (2006) The small GTPase R-Ras regulates organization of actin and drives membrane protrusions through the activity of PLCepsilon. J Cell Sci 119(Pt 7): 1307-19.
- Giannone G, Sheetz MP (2006) Substrate rigidity and force define form through tyrosine phosphatase and kinase pathways Trends Cell Biol 16(4): 213-23.
- Jiang G, Huang AH, Cai Y, Tanase M, Sheetz MP (2006) Rigidity sensing at the leading edge through alphavbeta3 integrins and RPTPalpha. Biophys J 90(5): 1804-9.
- von Wichert G, Sheetz MP. (2005) Mechanisms of disease: the biophysical interpretation of the ECM affects physiological and pathophysiological cellular behavior. Z Gastroenterol 43(12): 1329-36.
- De Vos KJ, Allan VJ, Grierson AJ, Sheetz MP (2005) Mitochondrial function and actin regulate dynamin-related protein 1-dependent mitochondrial fission. Curr Biol 15(7): 678-83.
- Miller LW, Cai Y, Sheetz MP, Cornish VW. (2005) In vivo protein labeling with trimethoprim conjugates: a flexible chemical tag. Nat Methods 2(4): 255-7.
- Döbereiner HG, Dubin-Thaler BJ, Giannone G, Sheetz MP. (2005) Force sensing and generation in cell phases: analyses of complex functions. J Appl Physiol 98(4): 1542-6.
- Meshel AS, Wei Q, Adelstein RS, Sheetz MP. (2005) Basic mechanism of three-dimensional collagen fibre transport by fibroblasts. Nat Cell Biol 7(2): 157-64.
- Tamada M, Sheetz MP, Sawada Y. (2004) Activation of a signaling cascade by cytoskeleton stretch Dev Cell 2004: 709-18.
- Döbereiner HG, Dubin-Thaler B, Giannone G, Xenias HS, Sheetz MP. (2004) Dynamic phase transitions in cell spreading.Phys Rev Lett 93(10): 108105.
- Maier B, Koomey M, Sheetz MP (2004) A force-dependent switch reverses type IV pilus retraction. Proc Natl Acad Sci101(30): 10961-6.
- De Vos, K., J. Sable, K.E. Miller, and M.P. Sheetz (2003) Expression of phosphatidylinositol (4.5) bisphosphate-specific pleckstrin homology domains alters direction but not level of axonal transport of mitochondria Mol Biol Cell In Press.
- Jiang, G., G. Giannone, D.R. Critchley, E. Fukumoto, and M.P. Sheetz (2003) Two-piconewton slip bond between fibronectin and the cytoskeleton depends on talin Nature 424: 334-7.Article
- Galbraith, C.G., K.M. Yamada, and M.P. Sheetz (2002) The relationship between force and focal complex development J Cell Biol 159: 695-705. Article
- Maier, B., L. Potter, M. So, C.D. Long, H.S. Seifert, and M.P. Sheetz (2002) Single pilus motor forces exceed 100 pN Proc Natl Acad Sci U S A 99: 16012-7. Article
- Sawada, Y., and M.P. Sheetz (2002) Force transduction by Triton cytoskeletons J Cell Biol 156: 609-15. Article
- Sheetz, M.P. (2001) Cell control by membrane-cytoskeleton adhesion Nat. Rev. Molec. Cell Biol. 2: 392-396. Article
- Merz, A. J., M. So, and M.P. Sheetz. (2000) Pilus retraction powers bacterial twitching motility Nature 407: 98-102.Article
- Miller, K. E. and M.P. Sheetz (2000) Characterization of myosin V binding to brain vesicles J. Biol. Chem. 275: 2598-2606.
- Raucher, D., and M. P. Sheetz. (2000) Cell spreading is regulated by membrane tension J. Cell Biol. 148: 127-136.Article
- Raucher, D., T. Stauffer, W. Chen, K. Shen, S. Guo, J.D. York, M.P. Sheetz, and T. Meyer (2000) Phosphatidylinositol 4,5-bisphosphate functions as a second messenger that regulates cytoskeleton-plasma membrane adhesion Cell 100: 221-228.Article
- Felsenfeld, D.P., Schwartzberg, P. L., Venegas, A., Tse, R. and Sheetz, M.P. (1999) Selective regulation of integrin-cytoskeleton interactions by the tyrosine kinase Src Nature Cell Biol. 1: 200-206.
- Galbraith, C. G. and M.P. Sheetz (1999) Keratocytes pull with similar forces on their dorsal and ventral surfaces J. Cell Biol.147: 1313-1323.
- Raucher, D., and Sheetz, M. P. (1999) Membrane expansion increases endocytosis rate during mitosis J. Cell Biol. 144: 497-506.
- Choquet, D., D.P. Felsenfeld, and M.P. Sheetz. (1997) Extracellular Matrix Rigidity Causes Strengthening of Integrin-Cytoskeletal Linkages Cell 88: 39-48.