Elucidation of sigma factor-associated networks in Pseudomonas aeruginosa reveals a modular architecture with limited and function-specific crosstalk

PLoS Pathog. 2015 Mar 17;11(3):e1004744. doi: 10.1371/journal.ppat.1004744. eCollection 2015 Mar.

Abstract

Sigma factors are essential global regulators of transcription initiation in bacteria which confer promoter recognition specificity to the RNA polymerase core enzyme. They provide effective mechanisms for simultaneously regulating expression of large numbers of genes in response to challenging conditions, and their presence has been linked to bacterial virulence and pathogenicity. In this study, we constructed nine his-tagged sigma factor expressing and/or deletion mutant strains in the opportunistic pathogen Pseudomonas aeruginosa. To uncover the direct and indirect sigma factor regulons, we performed mRNA profiling, as well as chromatin immunoprecipitation coupled to high-throughput sequencing. We furthermore elucidated the de novo binding motif of each sigma factor, and validated the RNA- and ChIP-seq results by global motif searches in the proximity of transcriptional start sites (TSS). Our integrated approach revealed a highly modular network architecture which is composed of insulated functional sigma factor modules. Analysis of the interconnectivity of the various sigma factor networks uncovered a limited, but highly function-specific, crosstalk which orchestrates complex cellular processes. Our data indicate that the modular structure of sigma factor networks enables P. aeruginosa to function adequately in its environment and at the same time is exploited to build up higher-level functions by specific interconnections that are dominated by a participation of RpoN.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin Immunoprecipitation
  • Cluster Analysis
  • High-Throughput Nucleotide Sequencing
  • Host-Parasite Interactions / physiology*
  • Pseudomonas Infections / genetics*
  • Pseudomonas Infections / metabolism
  • Pseudomonas aeruginosa / physiology*
  • Receptor Cross-Talk / physiology
  • Sequence Analysis, RNA
  • Sigma Factor / genetics*
  • Sigma Factor / metabolism
  • Signal Transduction / physiology*
  • Transcriptome

Substances

  • Sigma Factor

Associated data

  • GEO/GSE54997
  • GEO/GSE54998
  • GEO/GSE54999

Grants and funding

SH was supported by an ERC starter grant (RESISTOME 260276), the German Research Foundation (DFG SFB 900) and the Federal Ministry of Education and Research (1616038C). SS and SB were supported by the President`s Initiative and Networking Fund of the Helmholtz Association of German Research Centers (HGF) under contract number VH-GS-202. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.