Synthetic Cystic Fibrosis Sputum Medium Regulates Flagellar Biosynthesis Through the flhF Gene in Burkholderia Cenocepacia

Front Cell Infect Microbiol. 2016 Jun 14;6:65. doi: 10.3389/fcimb.2016.00065. eCollection 2016.

Abstract

Burkholderia cenocepacia belongs to the Burkholderia cepacia complex (Bcc), a group of at least 18 distinct species that establish chronic infections in the lung of people with the genetic disease cystic fibrosis (CF). The sputum of CF patients is rich in amino acids and was previously shown to increase flagellar gene expression in B. cenocepacia. We examined flagellin expression and flagellar morphology of B. cenocepacia grown in synthetic cystic fibrosis sputum medium (SCFM) compared to minimal medium. We found that CF nutritional conditions induce increased motility and flagellin expression. Individual amino acids added at the same concentrations as found in SCFM also increased motility but not flagellin expression, suggesting a chemotactic effect of amino acids. Electron microscopy and flagella staining demonstrated that the increase in flagellin corresponds to a change in the number of flagella per cell. In minimal medium, the ratio of multiple: single: aflagellated cells was 2:3.5:4.5; while under SCFM conditions, the ratio was 7:2:1. We created a deletion mutant, ΔflhF, to study whether this putative GTPase regulates the flagellation pattern of B. cenocepacia K56-2 during growth in CF conditions. The ΔflhF mutant exhibited 80% aflagellated, 14% single and 6% multiple flagellated bacterial subpopulations. Moreover, the ratio of multiple to single flagella in WT and ΔflhF was 3.5 and 0.43, respectively in CF conditions. The observed differences suggest that FlhF positively regulates flagellin expression and the flagellation pattern in B. cenocepacia K56-2 during CF nutritional conditions.

Keywords: Burkholderia cenocepacia; SCFM; cystic fibrosis; flagellum; flhF; motility.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / metabolism
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Burkholderia Infections / microbiology
  • Burkholderia cenocepacia / genetics*
  • Burkholderia cenocepacia / growth & development
  • Burkholderia cenocepacia / metabolism*
  • Burkholderia cenocepacia / pathogenicity
  • Burkholderia cepacia complex / classification
  • Cell Migration Assays
  • Cystic Fibrosis*
  • Escherichia coli / genetics
  • Flagella / chemistry
  • Flagella / genetics
  • Flagellin / biosynthesis*
  • Flagellin / genetics*
  • Gene Expression Regulation, Bacterial
  • Microscopy, Electron
  • Monomeric GTP-Binding Proteins / genetics*
  • Monomeric GTP-Binding Proteins / metabolism
  • Phenotype
  • Sequence Deletion
  • Sputum / chemistry*

Substances

  • Amino Acids
  • Bacterial Proteins
  • Flagellin
  • flhF protein, Bacteria
  • Monomeric GTP-Binding Proteins