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Mapping Local and Global Liquid Phase Behavior in Living Cells Using Photo-Oligomerizable Seeds

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Bracha, Dan, Walls, Mackenzie T., Wei, Ming-Tzo, Zhu, Lian, Kurian, Martin, Avalos, José L., Toettcher, Jared E., Brangwynne, Clifford P.·2018·Cell
DOI 10.1016/j.cell.2018.10.048PMC PMC6724719

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Abstract

Liquid-liquid phase separation plays a key role in the assembly of diverse intracellular structures. However, the biophysical principles by which phase separation can be precisely localized within subregions of the cell are still largely unclear, particularly for low-abundance proteins. Here, we introduce an oligomerizing biomimetic system, "Corelets," and utilize its rapid and quantitative light-controlled tunability to map full intracellular phase diagrams, which dictate the concentrations at which phase separation occurs and the transition mechanism, in a protein sequence dependent manner. Surprisingly, both experiments and simulations show that while intracellular concentrations may be insufficient for global phase separation, sequestering protein ligands to slowly diffusing nucleation centers can move the cell into a different region of the phase diagram, resulting in localized phase separation. This diffusive capture mechanism liberates the cell from the constraints of global protein abundance and is likely exploited to pattern condensates associated with diverse biological processes. VIDEO ABSTRACT.

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  • The enzymatic reaction is fast enough to overcome passive coarsening and slow enough to avoid spinodal decomposition.

    “We can thus ensure that the reaction is fast enough to overcome passive coarsening and slow enough to avoid spinodal decomposition19,20.”

    From Active coacervate droplets are protocells that grow and resist Ostwald ripening (2021) · cites this paper as [20]

    The Results section on FUSN Corelet phase diagrams describes coarsening and spinodal decomposition under different concentration conditions, but does not link either to an enzymatic reaction rate. Searches for reaction-rate constraints and passive coarsening found no passage establishing the claimed fast-enough and slow-enough bounds. The indexed passages provide no page numbers. (LLM, conf=0.74)

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Authors
Bracha, D., Walls, M. T., Wei, M.-T., Zhu, L., Kurian, M., Avalos, J. L., Toettcher, J. E., & Brangwynne, C. P.
Affiliations
  • Princeton University (US)
  • Howard Hughes Medical Institute (US)
Corresponding author
Brangwynne, Clifford P.
ORCID
Venue
Cell
Publisher
Elsevier BV
Published
2018-11
Volume
175
Issue
6
Pages
1467-1480.e13
ISSN
0092-8674 (ISSN-L)
DOI
10.1016/j.cell.2018.10.048
PMC
PMC6724719
OpenAlex
W2902026154
Cited by
462
References
2
Citations by year
2026: 45 · 2025: 64 · 2024: 61 · 2023: 55 · 2022: 70 · 2021: 77 · 2020: 44 · 2019: 42 · 2018: 4
FWCI
14.85
Type
journal article
Language
en
Update notices
erratum (2019-01-10)
Open access
Licence
Locations
  • Cell (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • Howard Hughes Medical Institute
  • NIH 4D Nucleome Program (U01 DA040601)
  • DARPA (HR0011-17-2-0010)
  • Princeton Center for Complex Materials
  • NSF
  • MRSEC (DMR 1420541)
  • NSF CAREER (1253035, CBET-1751840)
  • US-Israel Binational Science Foundation (2016508)
  • Human Frontiers Science Program (LT001447/2017-C)
  • Pew Charitable Trusts
  • National Science Foundation Graduate Research Fellowship Program (DGE-1656466)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
Keywords
liquid-liquid phase separation, biomolecular condensates, nucleation centers, low-abundance proteins, intracellular diffusion
MeSH terms
Animals, Cytoplasm, HeLa Cells, Humans, Microscopy, Fluorescence, Biomimetic Materials, NIH 3T3 Cells, Mice, HEK293 Cells

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