Phenotype overlap in Xylella fastidiosa is controlled by the cyclic di-GMP phosphodiesterase Eal in response to antibiotic exposure and diffusible signal factor-mediated cell-cell signaling

Appl Environ Microbiol. 2013 Jun;79(11):3444-54. doi: 10.1128/AEM.03834-12. Epub 2013 Mar 29.

Abstract

Eal is an EAL domain protein in Xylella fastidiosa homologous to one involved in resistance to tobramycin in Pseudomonas aeruginosa. EAL and HD-GYP domain proteins are implicated in the hydrolysis of the secondary messenger bis-(3'-5')-cyclic dimeric GMP (cyclic di-GMP). Cell density-dependent communication mediated by a Diffusible Signal Factor (DSF) also modulates cyclic di-GMP levels in X. fastidiosa, thereby controlling the expression of virulence genes and genes involved in insect transmission. The possible linkage of Eal to both extrinsic factors such as antibiotics and intrinsic factors such as quorum sensing, and whether both affect virulence, was thus addressed. Expression of eal was induced by subinhibitory concentrations of tobramycin, and an eal deletion mutant was more susceptible to this antibiotic than the wild-type strain and exhibited phenotypes similar to those of an rpfF deletion mutant blocked in DSF production, such as hypermotility, reduced biofilm formation, and hypervirulence to grape. Consistent with that, the rpfF mutant was more susceptible than the wild-type strain to tobramycin. Therefore, we propose that cell-cell communication and antibiotic stress can apparently lead to similar modulations of cyclic di-GMP in X. fastidiosa, resulting in similar phenotypes. However, the effect of cell density is dominant compared to that of antibiotic stress, since eal is suppressed by RpfF, which may prevent inappropriate behavioral changes in response to antibiotic stress when DSF accumulates.

Publication types

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

MeSH terms

  • 3',5'-Cyclic-GMP Phosphodiesterases / metabolism*
  • Amino Acid Sequence
  • Anti-Bacterial Agents / pharmacology*
  • Benzothiazoles
  • Biofilms / drug effects
  • Biofilms / growth & development
  • Cell Communication / physiology*
  • DNA Primers / genetics
  • Diamines
  • Drug Resistance / genetics
  • Escherichia coli
  • Gene Deletion
  • Genetic Complementation Test
  • Microbial Sensitivity Tests
  • Microscopy, Electron, Scanning
  • Molecular Sequence Data
  • Organic Chemicals
  • Phenotype*
  • Pseudomonas aeruginosa / enzymology
  • Quinolines
  • Sequence Alignment
  • Tobramycin / pharmacology
  • Vitis / microbiology
  • Xylella / drug effects
  • Xylella / enzymology*
  • Xylella / pathogenicity*
  • Xylella / physiology

Substances

  • Anti-Bacterial Agents
  • Benzothiazoles
  • DNA Primers
  • Diamines
  • Organic Chemicals
  • Quinolines
  • SYBR Green I
  • 3',5'-Cyclic-GMP Phosphodiesterases
  • Tobramycin