Abstract
Cells that grow in confined spaces eventually build up mechanical compressive stress. This growth-induced pressure decreases cell growth. Growth-induced pressure is important in a multitude of contexts, including cancer1,2,3, microbial infections4 and biofouling5; yet, our understanding of its origin and molecular consequences remains limited. Here we combine microfluidic confinement of the yeast Saccharomyces cerevisiae6 with rheological measurements using genetically encoded multimeric nanoparticles7 to reveal that growth-induced pressure is accompanied with an increase in a key cellular physical property: macromolecular crowding. We develop a fully calibrated model that predicts how increased macromolecular crowding hinders protein expression and thus diminishes cell growth. This model is sufficient to explain the coupling of growth rate to pressure without the need for specific molecular sensors or signalling cascades. As molecular crowding is similar across all domains of life, this could be a deeply conserved mechanism of biomechanical feedback that allows environmental sensing originating from the fundamental physical properties of cells.
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Data availability
Source data are available for this paper. All other data that support the plots within this paper and other findings of this study are available from the corresponding authors upon reasonable request.
Code availability
All the codes used in this paper are available from the corresponding authors upon reasonable request.
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Acknowledgements
We thank E. Kassianidou for the initial help with the laser ablation experiments. We thank A. Liang, C. Petzold and K. Dancel-Manning at the NYULH DART Microscopy Laboratory for consultation and assistance with the transmission electron microscopy work; this core is partially funded by the NYU Cancer Center Support Grant NIH/NCI P30CA016087. The technological realizations and associated research works were partly supported by the French RENATECH network (M.D.). L.J.H. was funded by NIH grants R01 GM132447 and R37 CA240765, the American Cancer Society, the Pershing Square Sohn Cancer Research Award and Chan Zuckerberg Initiative. We thank E. Rojas for fruitful discussions. L.J.H. and M.D. thank the FACE foundation for travel support.
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B.A. and M.D. designed and performed the experiments and data analysis. C.F.-D. and E.D. performed the AFM experiments. L.J.H. designed the strains used in the study. B.A., L.J.H. and M.D. wrote the manuscript.
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Extended data
Extended Data Fig. 1 Device used in the microfluidic experiments.
a. Cells are loaded in a culture chamber connected on its sides to narrow channels that are used to set the chemical environment. Confined growth lead to the buildup of GIP, which is measured through the deformation of the elastic chamber. b. The culture chamber is, similar to the device presented in a., connected to a set of narrow channels to set the chemical environment. A valve is actuated to confine the cell population and allow it to build up GIP. We estimate GIP by measuring the deformation of the PDMS membrane. Opening of the valve leads to a relaxation of GIP.
Extended Data Fig. 2 Impact of osmotic shock.
Ratio of nucleus and cytoplasm volume under osmotic shock and growth induced pressure.
Extended Data Fig. 3 Linear and exponential fits on the diffusion as a function of GIP data.
The score for each fit is presented. We superimposed the prediction of diffusion as a function of GIP for the 40nm-GEMs, as well as the corresponding score.
Extended Data Fig. 4 40nm-GEMs diffusion as a function of cell volume.
Model fit (Eq. (13)) of the experimental data to extract \(\xi _{40} = 7.4 \pm 2.5\) (r2 = 0.99).
Extended Data Fig. 5 Contribution of density and chamber volume change in the growth rate.
For simplicity, we denoted kg as the growth rate, \(k_\rho = \partial _t\rho /\rho\) as the contribution of cell density ρ to growth rate, and \(k_V = \partial _PV/V\;\partial _tP\) the contribution of the volume of the chamber V.
Extended Data Fig. 6 Induction time of PADH2-mCherry.
The induction time is plotted as a function of time. Inset: induction time plotted as a function of growth rate.
Extended Data Fig. 7 Protein production rate as a function of GIP.
In orange: data from the PADH2-mCherry promoter. In blue: data from the PHIS3-GFP promoter.
Extended Data Fig. 8 Laser ablation experiments and calibration of turgor pressure.
a. We used a high-intensity laser pulse to make a hole in a cell, forcing its deflation. The cell radius changes as a function of turgor pressure, cell wall elasticity, and thickness of the cell wall. b. Cells were punctured with a laser, resulting in a decrease in cell radius proportional to turgor pressure. The similar decrease in radius of WT and hog1Δ cells indicates that, absent osmotic perturbation, these cells develop similar amounts of turgor pressure. The decrease in radius of osmotically compressed (c = 1 M sorbitol) hog1Δ cells indicates that these cells are still pressurized, albeit to a reduced extent. c. We used transmission electron microscopy to measure the cell wall thickness. d. We performed AFM experiments, using small deformations (below 0.2 μm) to extract the effective elasticity of the cell. This elasticity provided a mathematical function of turgor pressure, cell wall elasticity, and cell wall thickness.
Extended Data Fig. 9 Growth induced pressure as a function of time.
In blue, for the WT cells. In orange, for the hog1Δ cells.
Extended Data Fig. 10 Measurement of the fluorescence intensity in the center versus the edge of the chamber prior to induction of PADH2-mCherry.
The data shows an insignificant (p-value = 0.16) 2.3% difference.
Supplementary information
Supplementary Information
Supplementary information containing a table of strains used along with the mathematical modelling.
Source data
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Alric, B., Formosa-Dague, C., Dague, E. et al. Macromolecular crowding limits growth under pressure. Nat. Phys. 18, 411–416 (2022). https://doi.org/10.1038/s41567-022-01506-1
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DOI: https://doi.org/10.1038/s41567-022-01506-1
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