Clusters of bacterial RNA polymerase are biomolecular condensates that assemble through liquid–liquid phase separation
| العنوان: | Clusters of bacterial RNA polymerase are biomolecular condensates that assemble through liquid–liquid phase separation |
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| المؤلفون: | David E. Cohn, Stephanie C. Weber, Baljyot Parmar, Samantha Graydon Tope, Albright Kim, Rodrigo Reyes-Lamothe, Nicolas Soubry, Stefan Biedzinski, James Wall, Anne-Marie Ladouceur |
| المصدر: | Proceedings of the National Academy of Sciences of the United States of America |
| بيانات النشر: | National Academy of Sciences, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | DNA, Bacterial, liquid–liquid phase separation, Direct evidence, medicine.disease_cause, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, RNA polymerase, medicine, Cluster (physics), Escherichia coli, Nucleoid, A-DNA, Polymerase, 030304 developmental biology, spatial organization, 0303 health sciences, Multidisciplinary, biology, single-molecule tracking, Escherichia coli Proteins, 030302 biochemistry & molecular biology, Cell Biology, DNA-Directed RNA Polymerases, Biological Sciences, Single Molecule Imaging, Transcription antitermination, chemistry, Biophysics, biology.protein, bacteria, biomolecular condensate, Transcriptional Elongation Factors, 030217 neurology & neurosurgery, Cell Nucleolus, Macromolecule |
| الوصف: | Significance Bacterial cells are small and were long thought to have little to no internal structure. However, advances in microscopy have revealed that bacteria do indeed contain subcellular compartments. But how these compartments form has remained a mystery. Recent progress in larger, more complex eukaryotic cells has identified a novel mechanism for intracellular organization known as liquid–liquid phase separation. This process causes certain types of molecules to concentrate within distinct compartments inside the cell. Here, we demonstrate that the same process also occurs in bacteria. This work, together with a growing body of literature, suggests that liquid–liquid phase separation is a common mechanism for intracellular organization in both eukaryotic and prokaryotic cells. Once described as mere “bags of enzymes,” bacterial cells are in fact highly organized, with many macromolecules exhibiting nonuniform localization patterns. Yet the physical and biochemical mechanisms that govern this spatial heterogeneity remain largely unknown. Here, we identify liquid–liquid phase separation (LLPS) as a mechanism for organizing clusters of RNA polymerase (RNAP) in Escherichia coli. Using fluorescence imaging, we show that RNAP quickly transitions from a dispersed to clustered localization pattern as cells enter log phase in nutrient-rich media. RNAP clusters are sensitive to hexanediol, a chemical that dissolves liquid-like compartments in eukaryotic cells. In addition, we find that the transcription antitermination factor NusA forms droplets in vitro and in vivo, suggesting that it may nucleate RNAP clusters. Finally, we use single-molecule tracking to characterize the dynamics of cluster components. Our results indicate that RNAP and NusA molecules move inside clusters, with mobilities faster than a DNA locus but slower than bulk diffusion through the nucleoid. We conclude that RNAP clusters are biomolecular condensates that assemble through LLPS. This work provides direct evidence for LLPS in bacteria and demonstrates that this process can serve as a mechanism for intracellular organization in prokaryotes and eukaryotes alike. |
| اللغة: | English |
| تدمد: | 1091-6490 0027-8424 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::93ee1bac0b9403402239312388b26ee5 http://europepmc.org/articles/PMC7414142 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....93ee1bac0b9403402239312388b26ee5 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
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