Quantitative reconstitution of yeast RNA processing bodies
Simon L Currie1,2, Wenmin Xing1,2, Denise Muhlrad3,4
1HHMI, UT Southwestern Medical Center, Dallas TX 75390.
Summary
Researchers reconstituted yeast RNA processing bodies (P bodies) using purified components. This study reveals that simple interactions between concentrated proteins and RNA can recapitulate the physical properties of these cellular condensates.
Area of Science:
- Cell Biology
- Biochemistry
- Biophysics
Background:
- Biomolecular condensates often form via liquid-liquid phase separation (LLPS).
- Reconstituting natural condensates is complex due to numerous components and varying cellular concentrations.
- Previous reconstitutions often lack quantitative cellular data and natural complexity.
Purpose of the Study:
- To quantitatively reconstitute yeast RNA processing bodies (P bodies) from purified components.
- To investigate the role of protein concentration and RNA in P body formation and dynamics.
- To determine if simple interactions can define the physical properties of cellular condensates.
Main Methods:
- Purification of seven highly concentrated P-body proteins from yeast.
- In vitro reconstitution of condensates using purified proteins and RNA at cellular concentrations.
- Analysis of condensate formation, protein partitioning, dynamics, and reversibility.
Main Results:
- Five of seven concentrated proteins formed homotypic condensates individually.
- Reconstituted P bodies at cellular concentrations showed reasonable agreement with cellular protein partitioning and dynamics.
- RNA influenced P body maturation, delaying it and promoting reversibility.
Conclusions:
- Quantitative reconstitution of P bodies is achievable using purified components.
- Simple interactions among concentrated proteins and RNA largely dictate condensate physical properties.
- This approach provides insights into the biophysics of intracellular phase separation.
