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| Title | Quantitative spatial analysis of chromatin biomolecular condensates using cryoelectron tomography. |
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| Journal, issue, pages | Proc Natl Acad Sci U S A, Vol. 122, Issue 19, Page e2426449122, Year 2025 |
| Publish date | May 13, 2025 |
Authors | Huabin Zhou / Joshua Hutchings / Momoko Shiozaki / Xiaowei Zhao / Lynda K Doolittle / Shixin Yang / Rui Yan / Nikki Jean / Margot Riggi / Zhiheng Yu / Elizabeth Villa / Michael K Rosen / ![]() |
| PubMed Abstract | Phase separation is an important mechanism to generate certain biomolecular condensates and organize the cell interior. Condensate formation and function remain incompletely understood due to ...Phase separation is an important mechanism to generate certain biomolecular condensates and organize the cell interior. Condensate formation and function remain incompletely understood due to difficulties in visualizing the condensate interior at high resolution. Here, we analyzed the structure of biochemically reconstituted chromatin condensates through cryoelectron tomography. We found that traditional blotting methods of sample preparation were inadequate, and high-pressure freezing plus focused ion beam milling was essential to maintain condensate integrity. To identify densely packed molecules within the condensate, we integrated deep learning-based segmentation with context-aware template matching. Our approaches were developed on chromatin condensates and were also effective on condensed regions of in situ native chromatin. Using these methods, we determined the average structure of nucleosomes to 6.1 and 12 Å resolution in reconstituted and native systems, respectively, found that nucleosomes form heterogeneous interaction networks in both cases, and gained insight into the molecular origins of surface tension in chromatin condensates. Our methods should be applicable to biomolecular condensates containing large and distinctive components in both biochemical reconstitutions and certain cellular systems. |
External links | Proc Natl Acad Sci U S A / PubMed:40327693 / PubMed Central |
| Methods | EM (subtomogram averaging) |
| Resolution | 6.1 - 22.0 Å |
| Structure data | ![]() EMDB-49923: Subtomogram average of nucleosome structure extracted from the HeLa cell nuclei ![]() EMDB-49924: Subtomogram averaging of nucleosomes in reconstituted chromatin condensates ![]() EMDB-49929: Subtomogram average of nucleosome from NIH3T3 cells, class 2 ![]() EMDB-53275: Subtomogram average of nucleosome from NIH3T3 cells, class 1 |
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