A cell’s ability to react to mechanical stimuli is known to be affected by the transport of transcription factors, the proteins responsible for regulating transcription of DNA into RNA, across the membrane enveloping its nucleus. Yet the molecular mechanisms by which mechanical cues control this process remain unclear. Here we show that one such protein, myocardin-related transcription factor A (MRTFA), is imported into the nucleus at a rate that is inversely correlated with its nanomechanical stability, but independent of its thermodynamic stability. Attaching mechanically stable proteins to MRTFA results in reduced gene expression and the subsequent slowing down of cell migration. We conclude that the mechanical unfolding of proteins regulates their nuclear translocation rate, and highlight the role of the nuclear pore complex as a selective mechanosensor that is capable of detecting forces as low as ∼10 pN. The modulation of the mechanical stability of transcription factors may represent a general strategy for the control of gene expression.
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Vogel, V. & Sheetz, M. Local force and geometry sensing regulate cell functions. Nat. Rev. Mol. Cell Biol. 7, 265–275 (2006).
Dupont, S. et al. Role of YAP/TAZ in mechanotransduction. Nature 474, 179–183 (2011).
Markiewicz, E. et al. The inner nuclear membrane protein emerin regulates beta-catenin activity by restricting its accumulation in the nucleus. EMBO J. 25, 3275–3285 (2006).
Moon, H. S., Even-Ram, S., Kleinman, H. K. & Cha, H. J. Zyxin is upregulated in the nucleus by thymosin beta4 in SiHa cells. Exp. Cell Res. 312, 3425–3431 (2006).
Fedorchak, G. R., Kaminski, A. & Lammerding, J. Cellular mechanosensing: getting to the nucleus of it all. Prog. Biophys. Mol. Biol. 115, 76–92 (2014).
Medjkane, S., Perez-Sanchez, C., Gaggioli, C., Sahai, E. & Treisman, R. Myocardin-related transcription factors and SRF are required for cytoskeletal dynamics and experimental metastasis. Nat. Cell Biol. 11, 257–268 (2009).
Olson, E. N. & Nordheim, A. Linking actin dynamics and gene transcription to drive cellular motile functions. Nat. Rev. Mol. Cell Biol. 11, 353–365 (2010).
Jain, N., Iyer, K. V., Kumar, A. & Shivashankar, G. V. Cell geometric constraints induce modular gene-expression patterns via redistribution of HDAC3 regulated by actomyosin contractility. Proc. Natl Acad. Sci. USA 110, 11349–11354 (2013).
Connelly, J. T. et al. Actin and serum response factor transduce physical cues from the microenvironment to regulate epidermal stem cell fate decisions. Nat. Cell Biol. 12, 711–718 (2010).
Miralles, F., Posern, G., Zaromytidou, A. I. & Treisman, R. Actin dynamics control SRF activity by regulation of its coactivator MAL. Cell 113, 329–342 (2003).
Ho, C. Y., Jaalouk, D. E., Vartiainen, M. K. & Lammerding, J. Lamin A/C and emerin regulate MKL1-SRF activity by modulating actin dynamics. Nature 497, 507–511 (2013).
Pawlowski, R., Rajakyla, E. K., Vartiainen, M. K. & Treisman, R. An actin-regulated importin alpha/beta-dependent extended bipartite NLS directs nuclear import of MRTF-A. EMBO J. 29, 3448–3458 (2010).
Mouilleron, S., Langer, C. A., Guettler, S., McDonald, N. Q. & Treisman, R. Structure of a pentavalent G-actin•MRTF-A complex reveals how G-actin controls nucleocytoplasmic shuttling of a transcriptional coactivator. Sci. Signal. 4, ra40 (2011).
Vartiainen, M. K., Guettler, S., Larijani, B. & Treisman, R. Nuclear actin regulates dynamic subcellular localization and activity of the SRF cofactor MAL. Science 316, 1749–1752 (2007).
Baarlink, C., Wang, H. & Grosse, R. Nuclear actin network assembly by formins regulates the SRF coactivator MAL. Science 340, 864–867 (2013).
Kim, S. J. et al. Integrative structure and functional anatomy of a nuclear pore complex. Nature 555, 475–482 (2018).
Maillard, R. A. et al. ClpX(P) generates mechanical force to unfold and translocate its protein substrates. Cell 145, 459–469 (2011).
Olivares, A. O., Kotamarthi, H. C., Stein, B. J., Sauer, R. T. & Baker, T. A. Effect of directional pulling on mechanical protein degradation by ATP-dependent proteolytic machines. Proc. Natl Acad. Sci. USA 4, ra40 (2011).
Rodriguez-Larrea, D. & Bayley, H. Multistep protein unfolding during nanopore translocation. Nat. Nanotechnol. 8, 288–295 (2013).
Maimon, T., Elad, N., Dahan, I. & Medalia, O. The human nuclear pore complex as revealed by cryo-electron tomography. Structure 20, 998–1006 (2012).
Lin, D. H. et al. Architecture of the symmetric core of the nuclear pore. Science 352, aaf1015 (2016).
Lemke, E. A. The multiple faces of disordered nucleoporins. J. Mol. Biol. 428, 2011–2024 (2016).
Grunwald, D. & Singer, R. H. Multiscale dynamics in nucleocytoplasmic transport. Curr. Opin. Cell Biol. 24, 100–106 (2012).
Schmidt, H. B. & Gorlich, D. Transport selectivity of nuclear pores, phase separation, and membraneless organelles. Trends Biochem. Sci. 41, 46–61 (2016).
Rout, M. P., Aitchison, J. D., Magnasco, M. O. & Chait, B. T. Virtual gating and nuclear transport: the hole picture. Trends Cell Biol. 13, 622–628 (2003).
Lim, R. Y. et al. Nanomechanical basis of selective gating by the nuclear pore complex. Science 318, 640–643 (2007).
Frey, S., Richter, R. P. & Gorlich, D. FG-rich repeats of nuclear pore proteins form a three-dimensional meshwork with hydrogel-like properties. Science 314, 815–817 (2006).
Lim, R. Y. et al. Flexible phenylalanine–glycine nucleoporins as entropic barriers to nucleocytoplasmic transport. Proc. Natl Acad. Sci. USA 103, 9512–9517 (2006).
Yamada, J. et al. A bimodal distribution of two distinct categories of intrinsically disordered structures with separate functions in FG nucleoporins. Mol. Cell Proteom. 9, 2205–2224 (2010).
Peters, R. Translocation through the nuclear pore complex: selectivity and speed by reduction-of-dimensionality. Traffic 6, 421–427 (2005).
Fisher, T. E., Oberhauser, A. F., Carrion-Vazquez, M., Marszalek, P. E. & Fernandez, J. M. The study of protein mechanics with the atomic force microscope. Trends Biochem. Sci. 24, 379–384 (1999).
Mehlin, H., Daneholt, B. & Skoglund, U. Translocation of a specific premessenger ribonucleoprotein particle through the nuclear pore studied with electron microscope tomography. Cell 69, 605–613 (1992).
Grunwald, D. & Singer, R. H. In vivo imaging of labelled endogenous beta-actin mRNA during nucleocytoplasmic transport. Nature 467, 604–607 (2010).
Stevens, B. J. & Swift, H. RNA transport from nucleus to cytoplasm in Chironomus salivary glands. J. Cell Biol. 31, 55–77 (1966).
Lowe, A. R. et al. Selectivity mechanism of the nuclear pore complex characterized by single cargo tracking. Nature 467, 600–603 (2010).
Elosegui-Artola, A. et al. Force triggers YAP nuclear entry by regulating transport across nuclear pores. Cell 171, 1397–1410 e1314 (2017).
Kudo, N. et al. Leptomycin B inhibition of signal-mediated nuclear export by direct binding to CRM1. Exp. Cell Res. 242, 540–547 (1998).
Li, H., Carrion-Vazquez, M., Oberhauser, A. F., Marszalek, P. E. & Fernandez, J. M. Point mutations alter the mechanical stability of immunoglobulin modules. Nat. Struct. Biol. 7, 1117–1120 (2000).
Kang, H. J., Coulibaly, F., Clow, F., Proft, T. & Baker, E. N. Stabilizing isopeptide bonds revealed in gram-positive bacterial pilus structure. Science 318, 1625–1628 (2007).
Alegre-Cebollada, J., Badilla, C. L. & Fernandez, J. M. Isopeptide bonds block the mechanical extension of pili in pathogenic Streptococcus pyogenes. J. Biol. Chem. 285, 11235–11242 (2010).
Kang, H. J. & Baker, E. N. Intramolecular isopeptide bonds give thermodynamic and proteolytic stability to the major pilin protein of Streptococcus pyogenes. J. Biol. Chem. 284, 20729–20737 (2009).
Li, H. et al. Reverse engineering of the giant muscle protein titin. Nature 418, 998–1002 (2002).
Li, H. & Fernandez, J. M. Mechanical design of the first proximal Ig domain of human cardiac titin revealed by single molecule force spectroscopy. J. Mol. Biol. 334, 75–86 (2003).
Carrion-Vazquez, M. et al. Mechanical and chemical unfolding of a single protein: a comparison. Proc. Natl Acad. Sci. USA 96, 3694–3699 (1999).
Randles, L. G., Rounsevell, R. W. & Clarke, J. Spectrin domains lose cooperativity in forced unfolding. Biophys. J. 92, 571–577 (2007).
Perez-Jimenez, R., Garcia-Manyes, S., Ainavarapu, S. R. & Fernandez, J. M. Mechanical unfolding pathways of the enhanced yellow fluorescent protein revealed by single molecule force spectroscopy. J. Biol. Chem. 281, 40010–40014 (2006).
Soderholm, J. F. et al. Importazole, a small molecule inhibitor of the transport receptor importin-beta. ACS Chem. Biol. 6, 700–708 (2011).
Niopek, D., Wehler, P., Roensch, J., Eils, R. & Di Ventura, B. Optogenetic control of nuclear protein export. Nat. Commun. 7, 10624 (2016).
Record, J. et al. Immunodeficiency and severe susceptibility to bacterial infection associated with a loss-of-function homozygous mutation of MKL1. Blood 126, 1527–1535 (2015).
Beck, M. & Hurt, E. The nuclear pore complex: understanding its function through structural insight. Nat. Rev. Mol. Cell Biol. 18, 73–89 (2017).
von Appen, A. et al. In situ structural analysis of the human nuclear pore complex. Nature 526, 140–143 (2015).
Bestembayeva, A. et al. Nanoscale stiffness topography reveals structure and mechanics of the transport barrier in intact nuclear pore complexes. Nat. Nanotechnol. 10, 60–64 (2015).
Frey, S. et al. Surface properties determining passage rates of proteins through nuclear pores. Cell 174, 202–217 e209 (2018).
Mohr, D., Frey, S., Fischer, T., Guttler, T. & Gorlich, D. Characterisation of the passive permeability barrier of nuclear pore complexes. EMBO J. 28, 2541–2553 (2009).
Ketterer, P. et al. DNA origami scaffold for studying intrinsically disordered proteins of the nuclear pore complex. Nat. Commun. 9, 902 (2018).
Popa, I., Kosuri, P., Alegre-Cebollada, J., Garcia-Manyes, S. & Fernandez, J. M. Force dependency of biochemical reactions measured by single-molecule force-clamp spectroscopy. Nat. Protoc. 8, 1261–1276 (2013).
Rosa, N. et al. Meltdown: a tool to help in the interpretation of thermal melt curves acquired by differential scanning fluorimetry. J. Biomol. Screen. 20, 898–905 (2015).
Rodriguez, L. G., Wu, X. & Guan, J. L. Wound-healing assay. Methods Mol. Biol. 294, 23–29 (2005).
We thank M. Vartiainen (University of Helsinki) for sharing the MRTFA–GFP plasmid, M. Parsons (King’s College London) for sharing the pEBFP2–N1 and pEYFP–N1 vectors and the MDA-MB-231 cell line, and U. Eggert and M. Pfuhl (King’s College London) for providing HeLa and Top10 competent cells, respectively. We thank the Nikon Imaging Centre at King’s College London for assistance in setting up the cell-imaging experiments. We wish to thank G. Yang for help in qPCR analysis, and C. Nichols and S. Conte (King’s College London) for help with differential scanning fluorimetry experiments. A.E.M.B. is the recipient of a Sir Henry Wellcome fellowship (210887/Z/18/Z). V.S.R. was funded by the BHF Centre for Research Excellence at King’s College London. This work was supported by BHF grant PG/13/50/30426, the European Commission (Mechanocontrol, grant agreement SEP-210342844), EPSRC Fellowship K00641X/1, EPSRC Strategic Equipment Grant EP/M022536/1, Leverhulme Trust Project Grant RPG-2015-225, Leverhulme Trust Research Leadership Award RL-2016-015, Wellcome Trust Investigator Award 212218/Z/18/Z and Royal Society Wolfson Fellowship RSWF/R3/183006, all to S.G.-M.
The authors declare no competing interests.
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Infante, E., Stannard, A., Board, S.J. et al. The mechanical stability of proteins regulates their translocation rate into the cell nucleus. Nat. Phys. 15, 973–981 (2019). https://doi.org/10.1038/s41567-019-0551-3
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