Diffusiophoresis promotes phase separation and transport of biomolecular condensates.


Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
03 Sep 2024
Historique:
received: 14 07 2023
accepted: 16 08 2024
medline: 4 9 2024
pubmed: 4 9 2024
entrez: 3 9 2024
Statut: epublish

Résumé

The internal microenvironment of a living cell is heterogeneous and comprises a multitude of organelles with distinct biochemistry. Amongst them are biomolecular condensates, which are membrane-less, phase-separated compartments enriched in system-specific proteins and nucleic acids. The heterogeneity of the cell engenders the presence of multiple spatiotemporal gradients in chemistry, charge, concentration, temperature, and pressure. Such thermodynamic gradients can lead to non-equilibrium driving forces for the formation and transport of biomolecular condensates. Here, we report how ion gradients impact the transport processes of biomolecular condensates on the mesoscale and biomolecules on the microscale. Utilizing a microfluidic platform, we demonstrate that the presence of ion concentration gradients can accelerate the transport of biomolecules, including nucleic acids and proteins, via diffusiophoresis. This hydrodynamic transport process allows localized enrichment of biomolecules, thereby promoting the location-specific formation of biomolecular condensates via phase separation. The ion gradients further impart directional motility of condensates, allowing them to exhibit enhanced diffusion along the gradient. Coupled with a reentrant phase behavior, the gradient-induced enhanced motility leads to a dynamical redistribution of condensates that ultimately extends their lifetime. Together, our results demonstrate diffusiophoresis as a non-equilibrium thermodynamic force that governs the formation and transport of biomolecular condensates.

Identifiants

pubmed: 39227569
doi: 10.1038/s41467-024-51840-6
pii: 10.1038/s41467-024-51840-6
doi:

Substances chimiques

Nucleic Acids 0
Proteins 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

7686

Subventions

Organisme : National Science Foundation (NSF)
ID : 2223737
Organisme : National Science Foundation (NSF)
ID : 2237177
Organisme : NIGMS NIH HHS
ID : R35 GM138186
Pays : United States

Informations de copyright

© 2024. The Author(s).

Références

Decker, C. J. & Parker, R. P-bodies and stress granules: possible roles in the control of translation and mRNA degradation. Cold Spring Harb. Perspect. Biol. 4, a012286 (2012).
pubmed: 22763747 pmcid: 3428773 doi: 10.1101/cshperspect.a012286
Su, X. et al. Phase separation of signaling molecules promotes T cell receptor signal transduction. Science 352, 595–599 (2016).
pubmed: 27056844 pmcid: 4892427 doi: 10.1126/science.aad9964
Nozaki, T. et al. Dynamic organization of chromatin domains revealed by super-resolution live-cell imaging. Mol. Cell 67, 282–293 (2017).
pubmed: 28712725 doi: 10.1016/j.molcel.2017.06.018
Pappu, R. V., Cohen, S. R., Dar, F., Farag, M. & Kar, M. Phase transitions of associative biomacromolecules. Chem. Rev. 123, 8945–8987 (2023).
Brangwynne, C. P., Tompa, P. & Pappu, R. V. Polymer physics of intracellular phase transitions. Nat. Phys. 11, 899–904 (2015).
doi: 10.1038/nphys3532
Brangwynne, C. P. et al. Germline P granules are liquid droplets that localize by controlled dissolution/condensation. Science 324, 1729–1732 (2009).
pubmed: 19460965 doi: 10.1126/science.1172046
Smith, J. et al. Spatial patterning of P granules by RNA-induced phase separation of the intrinsically-disordered protein MEG-3. eLife 5, e21337 (2016).
pubmed: 27914198 pmcid: 5262379 doi: 10.7554/eLife.21337
Lasker, K. et al. The material properties of a bacterial-derived biomolecular condensate tune biological function in natural and synthetic systems. Nat. Commun. 13, 5643 (2022).
pubmed: 36163138 pmcid: 9512792 doi: 10.1038/s41467-022-33221-z
Bowman, G. R. et al. Oligomerization and higher-order assembly contribute to sub-cellular localization of a bacterial scaffold: Mutational analysis of PopZ. Mol. Microbiol. 90, 776–795 (2013).
pubmed: 24102805 doi: 10.1111/mmi.12398
Powers, S. K. et al. Nucleo-cytoplasmic partitioning of ARF proteins controls auxin responses in Arabidopsis thaliana. Mol. Cell 76, 177–190 (2019).
pubmed: 31421981 pmcid: 6778021 doi: 10.1016/j.molcel.2019.06.044
Davis, R. B., Moosa, M. M. & Banerjee, P. R. Ectopic biomolecular phase transitions: fusion proteins in cancer pathologies. Trends Cell Biol. 32, 681–695 (2022).
pubmed: 35484036 pmcid: 9288518 doi: 10.1016/j.tcb.2022.03.005
Kiekebusch, D. & Thanbichler, M. Spatiotemporal organization of microbial cells by protein concentration gradients. Trends Microbiol. 22, 65–73 (2014).
pubmed: 24342487 doi: 10.1016/j.tim.2013.11.005
Keenan, T. M. & Folch, A. Biomolecular gradients in cell culture systems. Lab. Chip 8, 34–57 (2008).
pubmed: 18094760 doi: 10.1039/B711887B
King, M. R. et al. Macromolecular condensation organizes nucleolar sub-phases to set up a pH gradient. Cell 187, 1889–1906. (2024).
pubmed: 38503281 doi: 10.1016/j.cell.2024.02.029
Li, Y., Konstantopoulos, K., Zhao, R., Mori, Y. & Sun, S. X. The importance of water and hydraulic pressure in cell dynamics. J. Cell Sci. 133, jcs240341 (2020).
pubmed: 33087485 pmcid: 7595697 doi: 10.1242/jcs.240341
Albers, R. W. Biochemical aspects of active transport. Annu. Rev. Biochem. 36, 727–756 (1967).
pubmed: 18257736 doi: 10.1146/annurev.bi.36.070167.003455
Anderson, J. L. Colloid transport by interfacial forces. Annu. Rev. Fluid Mech 21, 61–99 (1989).
Shin, S. Diffusiophoretic separation of colloids in microfluidic flows. Phys. Fluids 32, 101302 (2020).
doi: 10.1063/5.0023415
Sear, R. P. Diffusiophoresis in cells: a general nonequilibrium, nonmotor mechanism for the metabolism-dependent transport of particles in cells. Phys. Rev. Lett. 122, 128101 (2019).
pubmed: 30978101 doi: 10.1103/PhysRevLett.122.128101
Chong, P. A., Vernon, R. M. & Forman-Kay, J. D. RGG/RG motif regions in RNA binding and phase separation. J. Mol. Biol. 430, 4650–4665 (2018).
pubmed: 29913160 doi: 10.1016/j.jmb.2018.06.014
Thandapani, P., O’Connor, T. R., Bailey, T. L. & Richard, S. Defining the RGG/RG motif. Mol. Cell 50, 613–623 (2013).
pubmed: 23746349 doi: 10.1016/j.molcel.2013.05.021
Alshareedah, I. et al. Interplay between short-range attraction and long-range repulsion controls reentrant liquid condensation of ribonucleoprotein–RNA complexes. J. Am. Chem. Soc. 141, 14593–14602 (2019).
pubmed: 31437398 pmcid: 7069731 doi: 10.1021/jacs.9b03689
Aumiller, W. M. & Keating, C. D. Phosphorylation-mediated RNA/peptide complex coacervation as a model for intracellular liquid organelles. Nat. Chem. 8, 129–137 (2016).
pubmed: 26791895 doi: 10.1038/nchem.2414
Boeynaems, S. et al. Spontaneous driving forces give rise to protein−RNA condensates with coexisting phases and complex material properties. Proc. Natl Acad. Sci. 116, 7889–7898 (2019).
pubmed: 30926670 pmcid: 6475405 doi: 10.1073/pnas.1821038116
Murthy, A. C. et al. Molecular interactions contributing to FUS SYGQ LC-RGG phase separation and co-partitioning with RNA polymerase II heptads. Nat. Struct. Mol. Biol. 28, 923–935 (2021).
pubmed: 34759379 pmcid: 8654040 doi: 10.1038/s41594-021-00677-4
Zhou, Q. et al. ATP regulates RNA‐driven cold inducible RNA binding protein phase separation. Protein Sci. 30, 1438–1453 (2021).
pubmed: 33991007 pmcid: 8197425 doi: 10.1002/pro.4123
Alshareedah, I., Moosa, M. M., Pham, M., Potoyan, D. A. & Banerjee, P. R. Programmable viscoelasticity in protein-RNA condensates with disordered sticker-spacer polypeptides. Nat. Commun. 12, 6620 (2021).
pubmed: 34785657 pmcid: 8595643 doi: 10.1038/s41467-021-26733-7
Banerjee, P. R., Milin, A. N., Moosa, M. M., Onuchic, P. L. & Deniz, A. A. Reentrant phase transition drives dynamic substructure formation in ribonucleoprotein droplets. Angew. Chem. Int. Ed. 56, 11354–11359 (2017).
doi: 10.1002/anie.201703191
Alshareedah, I., Moosa, M. M., Raju, M., Potoyan, D. A. & Banerjee, P. R. Phase transition of RNA−protein complexes into ordered hollow condensates. Proc. Natl Acad. Sci. 117, 15650–15658 (2020).
pubmed: 32571937 pmcid: 7354941 doi: 10.1073/pnas.1922365117
Alshareedah, I., Thurston, G. M. & Banerjee, P. R. Quantifying viscosity and surface tension of multicomponent protein-nucleic acid condensates. Biophys. J. 120, 1161–1169 (2021).
pubmed: 33453268 pmcid: 8059090 doi: 10.1016/j.bpj.2021.01.005
Paustian, J. S., Azevedo, R. N., Lundin, S.-T. B., Gilkey, M. J. & Squires, T. M. Microfluidic microdialysis: spatiotemporal control over solution microenvironments using integrated hydrogel membrane microwindows. Phys. Rev. X 3, 041010 (2013).
Shah, P. R. et al. Temperature dependence of diffusiophoresis via a novel microfluidic approach. Lab. Chip 22, 1980–1988 (2022).
pubmed: 35445222 doi: 10.1039/D1LC00916H
Jones, D. P. Intracellular diffusion gradients of O2 and ATP. Am. J. Physiol. -Cell Physiol. 250, C663–C675 (1986).
doi: 10.1152/ajpcell.1986.250.5.C663
Kaur, T. et al. Sequence-encoded and composition-dependent protein-RNA interactions control multiphasic condensate morphologies. Nat. Commun. 12, 872 (2021).
pubmed: 33558506 pmcid: 7870978 doi: 10.1038/s41467-021-21089-4
Shin, S. et al. Size-dependent control of colloid transport via solute gradients in dead-end channels. Proc. Natl Acad. Sci. 113, 257–261 (2016).
pubmed: 26715753 doi: 10.1073/pnas.1511484112
Shin, S., Ault, J. T., Feng, J., Warren, P. B. & Stone, H. A. Low-cost zeta potentiometry using solute gradients. Adv. Mater. 29, 1701516 (2017).
doi: 10.1002/adma.201701516
Doan, V. S., Chun, S., Feng, J. & Shin, S. Confinement-dependent diffusiophoretic transport of nanoparticles in collagen hydrogels. Nano Lett. 21, 7625–7630 (2021).
pubmed: 34516140 doi: 10.1021/acs.nanolett.1c02251
Doan, V. S., Kim, D.-O., Snoeyink, C., Sun, Y. & Shin, S. Shape- and orientation-dependent diffusiophoresis of colloidal ellipsoids. Phys. Rev. E 107, L052602 (2023).
pubmed: 37329064 doi: 10.1103/PhysRevE.107.L052602
Palacci, J., Cottin-Bizonne, C., Ybert, C. & Bocquet, L. Osmotic traps for colloids and macromolecules based on logarithmic sensing in salt taxis. Soft Matter 8, 980–994 (2012).
doi: 10.1039/C1SM06395B
Shin, S., Warren, P. B. & Stone, H. A. Cleaning by surfactant gradients: particulate removal from porous materials and the significance of rinsing in laundry detergency. Phys. Rev. Appl. 9, 034012 (2018).
doi: 10.1103/PhysRevApplied.9.034012
Peter, Q. A. E. et al. Microscale diffusiophoresis of proteins. J. Phys. Chem. B 126, 8913–8920 (2022).
pubmed: 36306420 pmcid: 9661530 doi: 10.1021/acs.jpcb.2c04029
Annunziata, O., Buzatu, D. & Albright, J. G. Protein diffusiophoresis and salt osmotic diffusion in aqueous solutions. J. Phys. Chem. B 116, 12694–12705 (2012).
pubmed: 23013156 doi: 10.1021/jp307625d
Doan, V. S., Saingam, P., Yan, T. & Shin, S. A trace amount of surfactants enables diffusiophoretic swimming of bacteria. ACS Nano 14, 14219–14227 (2020).
pubmed: 33000940 doi: 10.1021/acsnano.0c07502
Shin, S., Doan, V. S. & Feng, J. Osmotic delivery and release of lipid-encapsulated molecules via sequential solution exchange. Phys. Rev. Appl. 12, 024014 (2019).
doi: 10.1103/PhysRevApplied.12.024014
Rasmussen, M. K., Pedersen, J. N. & Marie, R. Size and surface charge characterization of nanoparticles with a salt gradient. Nat. Commun. 11, 2337 (2020).
pubmed: 32393750 pmcid: 7214416 doi: 10.1038/s41467-020-15889-3
Shin, S., Ault, J. T., Warren, P. B. & Stone, H. A. Accumulation of colloidal particles in flow junctions induced by fluid flow and diffusiophoresis. Phys. Rev. X 7, 041038 (2017).
Palacci, J., Abécassis, B., Cottin-Bizonne, C., Ybert, C. & Bocquet, L. Colloidal motility and pattern formation under rectified diffusiophoresis. Phys. Rev. Lett. 104, 138302 (2010).
pubmed: 20481918 doi: 10.1103/PhysRevLett.104.138302
Lafontaine, D. L. J., Riback, J. A., Bascetin, R. & Brangwynne, C. P. The nucleolus as a multiphase liquid condensate. Nat. Rev. Mol. Cell Biol. 22, 165–182 (2021).
pubmed: 32873929 doi: 10.1038/s41580-020-0272-6
Riback, J. A. et al. Viscoelasticity and advective flow of RNA underlies nucleolar form and function. Mol. Cell 83, 3095–3107 (2023).
Abécassis, B., Cottin-Bizonne, C., Ybert, C., Ajdari, A. & Bocquet, L. Boosting migration of large particles by solute contrasts. Nat. Mater. 7, 785–789 (2008).
pubmed: 18711384 doi: 10.1038/nmat2254
Raynal, F. & Volk, R. Diffusiophoresis, Batchelor scale and effective Péclet numbers. J. Fluid Mech. 876, 818–829 (2019).
doi: 10.1017/jfm.2019.589
Florea, D., Musa, S., Huyghe, J. M. R. & Wyss, H. M. Long-range repulsion of colloids driven by ion exchange and diffusiophoresis. Proc. Natl Acad. Sci. 111, 6554–6559 (2014).
pubmed: 24748113 pmcid: 4020040 doi: 10.1073/pnas.1322857111
Velegol, D., Garg, A., Guha, R., Kar, A. & Kumar, M. Origins of concentration gradients for diffusiophoresis. Soft Matter 12, 4686–4703 (2016).
pubmed: 27174044 doi: 10.1039/C6SM00052E
Jung, B., Bharadwaj, R. & Santiago, J. G. Thousandfold signal increase using field-amplified sample stacking for on-chip electrophoresis. Electrophoresis 24, 3476–3483 (2003).
pubmed: 14595694 doi: 10.1002/elps.200305611
Kar, M. et al. Phase-separating RNA-binding proteins form heterogeneous distributions of clusters in subsaturated solutions. Proc. Natl Acad. Sci. 119, e2202222119 (2022).
pubmed: 35787038 pmcid: 9282234 doi: 10.1073/pnas.2202222119
Fritsch, A. W. et al. Local thermodynamics govern formation and dissolution of Caenorhabditis elegans P granule condensates. Proc. Natl Acad. Sci. 118, e2102772118 (2021).
pubmed: 34507991 pmcid: 8449359 doi: 10.1073/pnas.2102772118
Chiang, T.-Y. & Velegol, D. Multi-ion diffusiophoresis. J. Colloid Interface Sci. 424, 120–123 (2014).
pubmed: 24767507 doi: 10.1016/j.jcis.2014.03.003
Gupta, A., Rallabandi, B. & Stone, H. A. Diffusiophoretic and diffusioosmotic velocities for mixtures of valence-asymmetric electrolytes. Phys. Rev. Fluids 4, 043702 (2019).
doi: 10.1103/PhysRevFluids.4.043702
Alshareedah, I. et al. Determinants of viscoelasticity and flow activation energy in biomolecular condensates. Sci. Adv. 10, eadi6539 (2024).
Young, N. O., Goldstein, J. S. & Block, M. J. The motion of bubbles in a vertical temperature gradient. J. Fluid Mech. 6, 350–356 (1959).
doi: 10.1017/S0022112059000684
Guo, W. et al. Non-associative phase separation in an evaporating droplet as a model for prebiotic compartmentalization. Nat. Commun. 12, 3194 (2021).
pubmed: 34045455 pmcid: 8160217 doi: 10.1038/s41467-021-23410-7
Ianeselli, A. et al. Non-equilibrium conditions inside rock pores drive fission, maintenance and selection of coacervate protocells. Nat. Chem. 14, 32–39 (2022).
pubmed: 34873298 doi: 10.1038/s41557-021-00830-y
Testa, A. et al. Sustained enzymatic activity and flow in crowded protein droplets. Nat. Commun. 12, 6293 (2021).
pubmed: 34725341 pmcid: 8560906 doi: 10.1038/s41467-021-26532-0
Demarchi, L., Goychuk, A., Maryshev, I. & Frey, E. Enzyme-enriched condensates show self-propulsion, positioning, and coexistence. Phys. Rev. Lett. 130, 128401 (2023).
pubmed: 37027840 doi: 10.1103/PhysRevLett.130.128401
Jambon-Puillet, E. et al. Phase-separated droplets swim to their dissolution. Nat. Commun. 15, 3919 (2024).
pubmed: 38724503 pmcid: 11082165 doi: 10.1038/s41467-024-47889-y
Agrawal, A., Ganai, N., Sengupta, S. & Menon, G. I. Nonequilibrium biophysical processes influence the large-scale architecture of the cell nucleus. Biophys. J. 118, 2229–2244 (2020).
pubmed: 31818465 doi: 10.1016/j.bpj.2019.11.017
Ramm, B. et al. A diffusiophoretic mechanism for ATP-driven transport without motor proteins. Nat. Phys. 17, 850–858 (2021).
doi: 10.1038/s41567-021-01213-3
Shim, S. et al. CO
pubmed: 33514979 doi: 10.1039/D0SM02023K
Vrhovec Hartman, S., Božič, B. & Derganc, J. Migration of blood cells and phospholipid vesicles induced by concentration gradients in microcavities. N. Biotechnol. 47, 60–66 (2018).
pubmed: 29501588 doi: 10.1016/j.nbt.2018.02.015
Hajian, R. & Hardt, S. Formation and lateral migration of nanodroplets via solvent shifting in a microfluidic device. Microfluid. Nanofluid. 19, 1281–1296 (2015).
doi: 10.1007/s10404-015-1644-7
Deamer, D. Concentration gradients. in Encyclopedia of Astrobiology (eds. Gargaud, M. et al.). https://doi.org/10.1007/978-3-642-11274-4_226 . 354–355 (Springer Berlin Heidelberg, 2011).
Bartolo, D., Degré, G., Nghe, P. & Studer, V. Microfluidic stickers. Lab. Chip 8, 274–279 (2008).
pubmed: 18231666 doi: 10.1039/B712368J
Nery-Azevedo, R., Banerjee, A. & Squires, T. M. Diffusiophoresis in ionic surfactant gradients. Langmuir 33, 9694–9702 (2017).
pubmed: 28853577 doi: 10.1021/acs.langmuir.7b01094
D’Orazio, T., Guaragnella, C., Leo, M. & Distante, A. A new algorithm for ball recognition using circle Hough transform and neural classifier. Pattern Recognit. 37, 393–408 (2004).
doi: 10.1016/S0031-3203(03)00228-0
Kirby, B. J. & Hasselbrink, E. F. Zeta potential of microfluidic substrates: 1. Theory, experimental techniques, and effects on separations. Electrophoresis 25, 187–202 (2004).
pubmed: 14743473 doi: 10.1002/elps.200305754

Auteurs

Viet Sang Doan (VS)

Department of Mechanical and Aerospace Engineering, University at Buffalo, The State University of New York, Buffalo, NY, USA.

Ibraheem Alshareedah (I)

Department of Physics, University at Buffalo, The State University of New York, Buffalo, NY, USA.

Anurag Singh (A)

Department of Physics, University at Buffalo, The State University of New York, Buffalo, NY, USA.

Priya R Banerjee (PR)

Department of Physics, University at Buffalo, The State University of New York, Buffalo, NY, USA. prbanerj@buffalo.edu.

Sangwoo Shin (S)

Department of Mechanical and Aerospace Engineering, University at Buffalo, The State University of New York, Buffalo, NY, USA. sangwoos@buffalo.edu.

Articles similaires

Databases, Protein Protein Domains Protein Folding Proteins Deep Learning
Adenosine Triphosphate Adenosine Diphosphate Mitochondrial ADP, ATP Translocases Binding Sites Mitochondria
Receptor, Cannabinoid, CB1 Ligands Molecular Dynamics Simulation Protein Binding Thermodynamics

Mutational analysis of Phanerochaete chrysosporium´s purine transporter.

Mariana Barraco-Vega, Manuel Sanguinetti, Gabriela da Rosa et al.
1.00
Phanerochaete Fungal Proteins Purines Aspergillus nidulans DNA Mutational Analysis

Classifications MeSH