Probing RNA Structure in Liquid-Liquid Phase Separation Using SHAPE-MaP

Methods Enzymol. 2018:611:67-79. doi: 10.1016/bs.mie.2018.09.039. Epub 2018 Nov 7.

Abstract

RNA is an integral component of many biological condensates. A variety of features of RNAs are linked to their function in biological phase separation. Length and negative charge provide fairly generic chemical inputs that drive condensation while sequence has been shown to influence both the molecular identity and biophysical properties of droplets. mRNA sequence guides the secondary structure of the polymers and RNA secondary structure licenses-specific RNA-RNA interactions and the recruitment of RNA-binding proteins. Here, we describe a method for directly probing the structure of mRNAs in the context of RNP-droplets formed via LLPS.

Keywords: Condensates; LLPS; Phase transitions; RNA; RNA structure.

MeSH terms

  • Animals
  • Escherichia coli / genetics
  • Gene Expression
  • Humans
  • Microscopy, Fluorescence / methods
  • Mutation
  • Nucleic Acid Conformation
  • Phase Transition*
  • RNA, Messenger / chemistry*
  • RNA, Messenger / genetics
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Ribonucleoproteins / chemistry
  • Ribonucleoproteins / genetics
  • Transcription, Genetic

Substances

  • RNA, Messenger
  • Recombinant Proteins
  • Ribonucleoproteins