Abstract
Liquid–liquid phase separation is increasingly recognized as a process involved in cellular organization. Thus far, a detailed structural characterization of this intrinsically heterogeneous process has been challenging. Here we combine solid- and solution-state NMR spectroscopy to obtain atomic-level insights into the assembly and maturation of cytoplasmic processing bodies that contain mRNA as well as enzymes involved in mRNA degradation. In detail, we have studied the enhancer of decapping 3 (Edc3) protein that is a central hub for processing body formation in yeast. Our results reveal that Edc3 domains exhibit diverse levels of structural organization and dynamics after liquid–liquid phase separation. In addition, we find that interactions between the different Edc3 domains and between Edc3 and RNA in solution are largely preserved in the condensed protein state, allowing processing bodies to rapidly form and dissociate upon small alterations in the cellular environment.
| Original language | English |
|---|---|
| Article number | 4536 |
| Journal | Nature Communications |
| Volume | 10 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 4 Oct 2019 |
Funding
This work was supported by the European Research Council (ERC) under the European Union’s Seventh Framework Programme (FP7/2007–2013), ERC Grant 616052 (RS). In addition, this study was funded by Netherlands Organization for Scientific Research (NWO) (grants 723.014.003 to M.W. and grants 700.26.121, 700.10.443, and 718.015.001 to M.B.) and iNEXT (project number 653706), a Horizon 2020 program of the European Union. Experiments at the 950 MHz instrument were supported by uNMR-NL, an NWO-funded Roadmap NMR Facility (no. 184.032.207). We thank David Meier and Olga Rudi for assisting in protein purification and Johanna Stöfl and Mira Schütz-Stoffregen for excellent technical assistance.
Keywords
- Intrinsically disordered proteins
- NMR spectroscopy
- Protein aggregation
- RNA