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Pré-Publication, Document De Travail Année : 2022

nucGEMs probe the biophysical properties of the nucleoplasm

Tamás Szórádi
Gururaj Kidiyoor
Ying Xie
Nora Herzog
Andrew Bazley
Martina Bonucci
Sarah Keegan
Farida Ettefa
Joël Lemiere
David Fenyö
Fred Chang
Liam Holt

Résumé

Abstract The cell interior is highly crowded and far from thermodynamic equilibrium. This environment can dramatically impact molecular motion and assembly, and therefore influence subcellular organization and biochemical reaction rates. These effects depend strongly on length-scale, with the least information available at the important mesoscale (10-100 nanometers), which corresponds to the size of crucial regulatory molecules such as RNA polymerase II. It has been challenging to study the mesoscale physical properties of the nucleoplasm because previous methods were labor-intensive and perturbative. Here, we report nuclear Genetically Encoded Multimeric nanoparticles (nucGEMs). Introduction of a single gene leads to continuous production and assembly of protein-based bright fluorescent nanoparticles of 40 nm diameter. We implemented nucGEMs in budding and fission yeast and in mammalian cell lines. We found differences in particle motility between the nucleus and the cytosol at the mesoscale, that mitotic chromosome condensation ejects nucGEMs from the nucleus, and that nucGEMs are excluded from heterochromatin and the nucleolus. nucGEMs enable hundreds of nuclear rheology experiments per hour, and allow evolutionary comparison of the physical properties of the cytosol and nucleoplasm.

Dates et versions

hal-03808263 , version 1 (10-10-2022)

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Tong Shu, Tamás Szórádi, Gururaj Kidiyoor, Ying Xie, Nora Herzog, et al.. nucGEMs probe the biophysical properties of the nucleoplasm. 2022. ⟨hal-03808263⟩
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