Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation

Khalil Joron, Juliane Oliveira Viegas, Liam Haas-Neill, Sariel Bier, Paz Drori, Shani Dvir, Patrick Siang Lin Lim, Sarah Rauscher, Eran Meshorer () and Eitan Lerner ()
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Khalil Joron: The Hebrew University of Jerusalem
Juliane Oliveira Viegas: The Hebrew University of Jerusalem, Edmond J. Safra Campus, Givat Ram
Liam Haas-Neill: University of Toronto Mississauga
Sariel Bier: The Hebrew University of Jerusalem
Paz Drori: The Hebrew University of Jerusalem
Shani Dvir: The Hebrew University of Jerusalem
Patrick Siang Lin Lim: The Hebrew University of Jerusalem, Edmond J. Safra Campus, Givat Ram
Sarah Rauscher: University of Toronto Mississauga
Eran Meshorer: The Hebrew University of Jerusalem, Edmond J. Safra Campus, Givat Ram
Eitan Lerner: The Hebrew University of Jerusalem

Nature Communications, 2023, vol. 14, issue 1, 1-18

Abstract: Abstract Fluorescent proteins (FP) are frequently used for studying proteins inside cells. In advanced fluorescence microscopy, FPs can report on additional intracellular variables. One variable is the local density near FPs, which can be useful in studying densities within cellular bio-condensates. Here, we show that a reduction in fluorescence lifetimes of common monomeric FPs reports increased levels of local densities. We demonstrate the use of this fluorescence-based variable to report the distribution of local densities within heterochromatin protein 1α (HP1α) in mouse embryonic stem cells (ESCs), before and after early differentiation. We find that local densities within HP1α condensates in pluripotent ESCs are heterogeneous and cannot be explained by a single liquid phase. Early differentiation, however, induces a change towards a more homogeneous distribution of local densities, which can be explained as a liquid-like phase. In conclusion, we provide a fluorescence-based method to report increased local densities and apply it to distinguish between homogeneous and heterogeneous local densities within bio-condensates.

Date: 2023
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DOI: 10.1038/s41467-023-40647-6

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