RNA is a critical element for the sizing and the composition of phase-separated RNA-protein condensates

Nat Commun. 2019 Jul 19;10(1):3230. doi: 10.1038/s41467-019-11241-6.

Abstract

Liquid-liquid phase separation is thought to be a key organizing principle in eukaryotic cells to generate highly concentrated dynamic assemblies, such as the RNP granules. Numerous in vitro approaches have validated this model, yet a missing aspect is to take into consideration the complex molecular mixture and promiscuous interactions found in vivo. Here we report the versatile scaffold ArtiG to generate concentration-dependent RNA-protein condensates within living cells, as a bottom-up approach to study the impact of co-segregated endogenous components on phase separation. We demonstrate that intracellular RNA seeds the nucleation of the condensates, as it provides molecular cues to locally coordinate the formation of endogenous high-order RNP assemblies. Interestingly, the co-segregation of intracellular components ultimately impacts the size of the phase-separated condensates. Thus, RNA arises as an architectural element that can influence the composition and the morphological outcome of the condensate phases in an intracellular context.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytoplasmic Granules / chemistry
  • Cytoplasmic Granules / metabolism*
  • HeLa Cells
  • Humans
  • Kinetics
  • Microscopy, Electron, Transmission
  • Protein Binding
  • Protein Interaction Maps
  • RNA / chemistry
  • RNA / metabolism*
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / metabolism*
  • Ribonucleoproteins / chemistry
  • Ribonucleoproteins / metabolism*
  • Ribonucleoproteins / ultrastructure

Substances

  • RNA-Binding Proteins
  • Ribonucleoproteins
  • RNA