How to find small non-coding RNAs in bacteria

Biol Chem. 2005 Dec;386(12):1219-38. doi: 10.1515/BC.2005.140.

Abstract

Small non-coding RNAs (sRNAs) have attracted considerable attention as an emerging class of gene expression regulators. In bacteria, a few regulatory RNA molecules have long been known, but the extent of their role in the cell was not fully appreciated until the recent discovery of hundreds of potential sRNA genes in the bacterium Escherichia coli. Orthologs of these E. coli sRNA genes, as well as unrelated sRNAs, were also found in other bacteria. Here we review the disparate experimental approaches used over the years to identify sRNA molecules and their genes in prokaryotes. These include genome-wide searches based on the biocomputational prediction of non-coding RNA genes, global detection of non-coding transcripts using microarrays, and shotgun cloning of small RNAs (RNomics). Other sRNAs were found by either co-purification with RNA-binding proteins, such as Hfq or CsrA/RsmA, or classical cloning of abundant small RNAs after size fractionation in polyacrylamide gels. In addition, bacterial genetics offers powerful tools that aid in the search for sRNAs that may play a critical role in the regulatory circuit of interest, for example, the response to stress or the adaptation to a change in nutrient availability. Many of the techniques discussed here have also been successfully applied to the discovery of eukaryotic and archaeal sRNAs.

Publication types

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

MeSH terms

  • Cloning, Molecular
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Eukaryotic Cells / metabolism
  • Gene Expression Regulation, Bacterial*
  • Genome, Bacterial
  • Host Factor 1 Protein / chemistry
  • Host Factor 1 Protein / genetics
  • Host Factor 1 Protein / metabolism
  • Oligonucleotide Array Sequence Analysis
  • RNA Processing, Post-Transcriptional
  • RNA, Archaeal / chemistry
  • RNA, Archaeal / genetics
  • RNA, Archaeal / metabolism
  • RNA, Bacterial* / chemistry
  • RNA, Bacterial* / genetics
  • RNA, Bacterial* / metabolism
  • RNA, Untranslated* / chemistry
  • RNA, Untranslated* / genetics
  • RNA, Untranslated* / metabolism
  • RNA-Binding Proteins* / chemistry
  • RNA-Binding Proteins* / genetics
  • RNA-Binding Proteins* / metabolism

Substances

  • Escherichia coli Proteins
  • Hfq protein, E coli
  • Host Factor 1 Protein
  • RNA, Archaeal
  • RNA, Bacterial
  • RNA, Untranslated
  • RNA-Binding Proteins