RNA-dependent proteome solubility maintenance in Escherichia coli lysates analysed by quantitative mass spectrometry: Proteomic characterization in terms of isoelectric point, structural disorder, functional hub, and chaperone network

RNA Biol. 2024 Jan;21(1):1-18. doi: 10.1080/15476286.2024.2315383. Epub 2024 Feb 15.

Abstract

Protein aggregation, a consequence of misfolding and impaired proteostasis, can lead to cellular malfunctions such as various proteinopathies. The mechanisms protecting proteins from aggregation in complex cellular environments have long been investigated, often from a protein-centric viewpoint. However, our study provides insights into a crucial, yet overlooked actor: RNA. We found that depleting RNAs from Escherichia coli lysates induces global protein aggregation. Our quantitative mass spectrometry analysis identified over 900 statistically significant proteins from the Escherichia coli proteome whose solubility depends on RNAs. Proteome-wide characterization showed that the RNA dependency is particularly enriched among acidic proteins, intrinsically disordered proteins, and structural hub proteins. Moreover, we observed distinct differences in RNA-binding mode and Gene Ontology categories between RNA-dependent acidic and basic proteins. Notably, the solubility of key molecular chaperones [Trigger factor, DnaJ, and GroES] is largely dependent on RNAs, suggesting a yet-to-be-explored hierarchical relationship between RNA-based chaperone (termed as chaperna) and protein-based chaperones, both of which constitute the whole chaperone network. These findings provide new insights into the RNA-centric role in maintaining healthy proteome solubility in vivo, where proteins associate with a variety of RNAs, either stably or transiently.

Keywords: Aggregation; RNA; RNA-binding proteins; intrinsically disordered regions (IDRs); molecular chaperones; proteostasis.

MeSH terms

  • Escherichia coli Proteins* / metabolism
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Isoelectric Point
  • Mass Spectrometry
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism
  • Protein Aggregates
  • Protein Folding
  • Proteome / metabolism
  • Proteomics
  • RNA / metabolism
  • Solubility

Substances

  • Proteome
  • RNA
  • Protein Aggregates
  • Escherichia coli Proteins
  • Molecular Chaperones