Quantitative spatial analysis of chromatin biomolecular condensates using cryoelectron tomography DOI Creative Commons
Huabin Zhou, Joshua Hutchings, Momoko Shiozaki

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(19)

Published: May 6, 2025

Phase separation is an important mechanism to generate certain biomolecular condensates and organize the cell interior. Condensate formation function remain incompletely understood due difficulties in visualizing condensate interior at high resolution. Here, we analyzed structure of biochemically reconstituted chromatin through cryoelectron tomography. We found that traditional blotting methods sample preparation were inadequate, high-pressure freezing plus focused ion beam milling was essential maintain integrity. To identify densely packed molecules within condensate, integrated deep learning–based segmentation with context-aware template matching. Our approaches developed on also effective condensed regions situ native chromatin. Using these methods, determined average nucleosomes 6.1 12 Å resolution systems, respectively, form heterogeneous interaction networks both cases, gained insight into molecular origins surface tension condensates. should be applicable containing large distinctive components biochemical reconstitutions cellular systems.

Language: Английский

Quantitative Spatial Analysis of Chromatin Biomolecular Condensates using Cryo-Electron Tomography DOI Creative Commons
Huabin Zhou, Joshua Hutchings, Momoko Shiozaki

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 2, 2024

Phase separation is an important mechanism to generate certain biomolecular condensates and organize the cell interior. Condensate formation function remain incompletely understood due difficulties in visualizing condensate interior at high resolution. Here we analyzed structure of biochemically reconstituted chromatin through cryo-electron tomography. We found that traditional blotting methods sample preparation were inadequate, high-pressure freezing plus focused ion beam milling was essential maintain integrity. To identify densely packed molecules within condensate, integrated deep learning-based segmentation with novel context-aware template matching. Our approaches developed on condensates, also effective condensed regions situ native chromatin. Using these methods, determined average nucleosomes 6.1 12 Å resolution systems, respectively, have a nearly random orientation distribution both cases. should be applicable diverse some cells.

Language: Английский

Citations

3
Quantitative spatial analysis of chromatin biomolecular condensates using cryoelectron tomography DOI Creative Commons
Huabin Zhou, Joshua Hutchings, Momoko Shiozaki

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(19)

Published: May 6, 2025

Phase separation is an important mechanism to generate certain biomolecular condensates and organize the cell interior. Condensate formation function remain incompletely understood due difficulties in visualizing condensate interior at high resolution. Here, we analyzed structure of biochemically reconstituted chromatin through cryoelectron tomography. We found that traditional blotting methods sample preparation were inadequate, high-pressure freezing plus focused ion beam milling was essential maintain integrity. To identify densely packed molecules within condensate, integrated deep learning–based segmentation with context-aware template matching. Our approaches developed on also effective condensed regions situ native chromatin. Using these methods, determined average nucleosomes 6.1 12 Å resolution systems, respectively, form heterogeneous interaction networks both cases, gained insight into molecular origins surface tension condensates. should be applicable containing large distinctive components biochemical reconstitutions cellular systems.

Language: Английский

Citations

0