| RRC ID |
66190
|
| 著者 |
Ladouceur AM, Parmar BS, Biedzinski S, Wall J, Tope SG, Cohn D, Kim A, Soubry N, Reyes-Lamothe R, Weber SC.
|
| タイトル |
Clusters of bacterial RNA polymerase are biomolecular condensates that assemble through liquid-liquid phase separation.
|
| ジャーナル |
Proc Natl Acad Sci U S A
|
| Abstract |
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.
|
| 巻・号 |
117(31)
|
| ページ |
18540-18549
|
| 公開日 |
2020-8-4
|
| DOI |
10.1073/pnas.2005019117
|
| PII |
2005019117
|
| PMID |
32675239
|
| PMC |
PMC7414142
|
| MeSH |
Cell Nucleolus / genetics
Cell Nucleolus / metabolism
DNA, Bacterial / genetics
DNA, Bacterial / metabolism
DNA-Directed RNA Polymerases / chemistry
DNA-Directed RNA Polymerases / genetics
DNA-Directed RNA Polymerases / metabolism*
Escherichia coli / chemistry
Escherichia coli / enzymology*
Escherichia coli / genetics
Escherichia coli / growth & development
Escherichia coli Proteins / genetics
Escherichia coli Proteins / metabolism
Single Molecule Imaging
Transcriptional Elongation Factors / genetics
Transcriptional Elongation Factors / metabolism
|
| IF |
9.412
|
| リソース情報 |
| 原核生物(大腸菌) |
ASKA library
JW3229
JW3138 |
