Techniques to Analyze sRNA Protein Cofactor Self-Assembly In Vitro

Methods Mol Biol. 2018:1737:321-340. doi: 10.1007/978-1-4939-7634-8_18.

Abstract

Post-transcriptional control of gene expression by small regulatory noncoding RNA (sRNA) needs protein accomplices to occur. Past research mainly focused on the RNA chaperone Hfq as cofactor. Nevertheless, recent studies indicated that other proteins might be involved in sRNA-based regulations. As some of these proteins have been shown to self-assemble, we describe in this chapter protocols to analyze the nano-assemblies formed. Precisely, we focus our analysis on Escherichia coli Hfq as a model, but the protocols presented here can be applied to analyze any polymer of proteins. This chapter thus provides a guideline to develop commonly used approaches to detect prokaryotic protein self-assembly, with a special focus on the detection of amyloidogenic polymers.

Keywords: Functional amyloid; Noncoding RNA cofactor; Protein self-assembly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Gene Expression Regulation, Bacterial
  • Host Factor 1 Protein / chemistry
  • Host Factor 1 Protein / genetics
  • Host Factor 1 Protein / metabolism*
  • In Vitro Techniques
  • Protein Binding
  • Protein Multimerization*
  • RNA, Bacterial / chemistry
  • RNA, Bacterial / genetics
  • RNA, Bacterial / metabolism*
  • RNA, Small Untranslated / chemistry
  • RNA, Small Untranslated / genetics
  • RNA, Small Untranslated / metabolism*

Substances

  • Escherichia coli Proteins
  • Hfq protein, E coli
  • Host Factor 1 Protein
  • RNA, Bacterial
  • RNA, Small Untranslated