Evaluating the Effect of Small RNAs and Associated Chaperones on Rho-Dependent Termination of Transcription In Vitro

Methods Mol Biol. 2018:1737:99-118. doi: 10.1007/978-1-4939-7634-8_7.

Abstract

Besides their well-known posttranscriptional effects on mRNA translation and decay, sRNAs and associated RNA chaperones (e.g., Hfq, CsrA) sometimes regulate gene expression at the transcriptional level. In this case, the sRNA-dependent machinery modulates the activity of the transcription termination factor Rho, a ring-shaped RNA translocase/helicase that dissociates transcription elongation complexes at specific loci of the bacterial genome. Here, we describe biochemical assays to detect Rho-dependent termination signals in genomic regions of interest and to assess the effects of sRNAs and/or associated RNA chaperones on such signals.

Keywords: Chaperone; Helicase; Hexamer; Rho; Ring-shaped; Termination; Transcription; sRNA.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Escherichia coli / genetics*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Gene Expression Regulation, Bacterial*
  • In Vitro Techniques
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism*
  • RNA, Bacterial / genetics
  • RNA, Small Untranslated / genetics*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Transcription Termination, Genetic*

Substances

  • Escherichia coli Proteins
  • Molecular Chaperones
  • RNA, Bacterial
  • RNA, Small Untranslated
  • RNA-Binding Proteins
  • Rho protein, E coli