A mechanism for MEX-5-driven disassembly of PGL-3/RNA condensates in vitro

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Natasha S. Lewis - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Silja Zedlitz - , Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck School Matter to Life (Author)
  • Hannes Ausserwöger - , University of Cambridge (Author)
  • Patrick M. McCall - , Clusters of Excellence PoL: Physics of Life, Chair of Spatiotemporal Organization of Subcellular Structures (PoL), Max Planck Institute of Molecular Cell Biology and Genetics, Center for Systems Biology Dresden (CSBD), Leibniz Institute of Polymer Research Dresden, Max-Planck-Institute for the Physics of Complex Systems (Author)
  • Lars Hubatsch - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Marco Nousch - , Martin Luther University Halle-Wittenberg (Author)
  • Martine Ruer-Gruß - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Carsten Hoege - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Frank Jülicher - , Max-Planck-Institute for the Physics of Complex Systems (Author)
  • Christian R. Eckmann - , Martin Luther University Halle-Wittenberg (Author)
  • Tuomas P.J. Knowles - , University of Cambridge (Author)
  • Anthony A. Hyman - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)

Abstract

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.

Details

Original languageEnglish
Article numbere2412218122
JournalProceedings of the National Academy of Sciences of the United States of America
Volume122
Issue number20
Publication statusPublished - 20 May 2025
Peer-reviewedYes

External IDs

PubMed 40354522

Keywords

ASJC Scopus subject areas

Keywords

  • C. elegans, phase-separation, polarity