The primary transcriptome of Neisseria meningitidis and its interaction with the RNA chaperone Hfq

Nucleic Acids Res. 2017 Jun 2;45(10):6147-6167. doi: 10.1093/nar/gkx168.

Abstract

Neisseria meningitidis is a human commensal that can also cause life-threatening meningitis and septicemia. Despite growing evidence for RNA-based regulation in meningococci, their transcriptome structure and output of regulatory small RNAs (sRNAs) are incompletely understood. Using dRNA-seq, we have mapped at single-nucleotide resolution the primary transcriptome of N. meningitidis strain 8013. Annotation of 1625 transcriptional start sites defines transcription units for most protein-coding genes but also reveals a paucity of classical σ70-type promoters, suggesting the existence of activators that compensate for the lack of -35 consensus sequences in N. meningitidis. The transcriptome maps also reveal 65 candidate sRNAs, a third of which were validated by northern blot analysis. Immunoprecipitation with the RNA chaperone Hfq drafts an unexpectedly large post-transcriptional regulatory network in this organism, comprising 23 sRNAs and hundreds of potential mRNA targets. Based on this data, using a newly developed gfp reporter system we validate an Hfq-dependent mRNA repression of the putative colonization factor PrpB by the two trans-acting sRNAs RcoF1/2. Our genome-wide RNA compendium will allow for a better understanding of meningococcal transcriptome organization and riboregulation with implications for colonization of the human nasopharynx.

MeSH terms

  • 3' Untranslated Regions / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Gene Expression Regulation, Bacterial*
  • Genes, Bacterial
  • Host Factor 1 Protein / metabolism*
  • MicroRNAs / classification
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Molecular Chaperones / metabolism*
  • Neisseria meningitidis / genetics*
  • Neisseria meningitidis / pathogenicity
  • Promoter Regions, Genetic
  • Protein Binding
  • RNA Stability
  • RNA, Bacterial / genetics*
  • RNA, Bacterial / metabolism
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Sequence Analysis, RNA
  • Species Specificity
  • Transcriptome*
  • Virulence

Substances

  • 3' Untranslated Regions
  • Bacterial Proteins
  • Host Factor 1 Protein
  • MicroRNAs
  • Molecular Chaperones
  • RNA, Bacterial
  • RNA, Messenger