A novel role for an ECF sigma factor in fatty acid biosynthesis and membrane fluidity in Pseudomonas aeruginosa

PLoS One. 2013 Dec 30;8(12):e84775. doi: 10.1371/journal.pone.0084775. eCollection 2013.

Abstract

Extracytoplasmic function (ECF) sigma factors are members of cell-surface signaling systems, abundant in the opportunistic pathogen Pseudomonas aeruginosa. Twenty genes coding for ECF sigma factors are present in P. aeruginosa sequenced genomes, most of them being part of TonB systems related to iron uptake. In this work, poorly characterized sigma factors were overexpressed in strain PA14, in an attempt to understand their role in the bacterium's physiology. Cultures overexpressing SigX displayed a biphasic growth curve, reaching stationary phase earlier than the control strain, followed by subsequent growth resumption. During the first stationary phase, most cells swell and die, but the remaining cells return to the wild type morphology and proceed to a second exponential growth. This is not due to compensatory mutations, since cells recovered from late time points and diluted into fresh medium repeated this behavior. Swollen cells have a more fluid membrane and contain higher amounts of shorter chain fatty acids. A proteomic analysis was performed to identify differentially expressed proteins due to overexpression of sigX, revealing the induction of several fatty acid synthesis (FAS) enzymes. Using qRT-PCR, we showed that at least one isoform from each of the FAS pathway enzymes were upregulated at the mRNA level in the SigX overexpressing strain thus pointing to a role for this ECF sigma factor in the FAS regulation in P. aeruginosa.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cell Membrane / genetics
  • Cell Membrane / metabolism*
  • Fatty Acids / biosynthesis*
  • Fatty Acids / genetics
  • Membrane Fluidity / physiology*
  • Pseudomonas aeruginosa / genetics
  • Pseudomonas aeruginosa / metabolism*
  • Sigma Factor / genetics
  • Sigma Factor / metabolism*

Substances

  • Bacterial Proteins
  • Fatty Acids
  • Sigma Factor

Grants and funding

This work was supported by grants 2009/09211-2 for RLB and 2008/03528-0 for ALB, São Paulo Research Foundation (FAPESP, www.fapesp.br) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, www.capes.gov.br). RLB was partially supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq,304349/2010-0, www.cnpq.br). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.