A mechanism for MEX-5-driven disassembly of PGL-3/RNA condensates in vitro
- Natasha S. Lewis
- Silja Zedlitz
- Hannes Ausserwöger
- Patrick M. McCall
- Lars Hubatsch
- Marco Nousch
- Martine Ruer-Gruß
- Carsten Hoege
- Frank Jülicher
- Christian R. Eckmann
- Tuomas P. J. Knowles
- Anthony A. Hyman
2025-05-12
MEX-5 regulates the formation and dissolution of P granules in Caenorhabditis elegans embryos, yet the thermodynamic basis of its activity remains unclear. Here, using a time-resolved in vitro reconstitution system, we show that MEX-5 dissolves preassembled liquid-like PGL-3/RNA condensates by altering RNA availability and shifting the phase boundary. We develop a microfluidic assay to systematically analyze how MEX-5 influences phase separation. By measuring the contribution of PGL-3 to phase separation, we show that MEX-5 reduces the free energy of PGL-3, shifting the equilibrium toward dissolution. Our findings provide a quantitative framework for understanding how RNA-binding proteins modulate condensate stability and demonstrate the power of microfluidics in precisely mapping phase transitions.