Using Light-Activated Enzymes for Modulating Intracellular c-di-GMP Levels in Bacteria

Methods Mol Biol. 2017:1657:169-186. doi: 10.1007/978-1-4939-7240-1_14.

Abstract

Signaling pathways involving second messenger c-di-GMP regulate various aspects of bacterial physiology and behavior. We describe the use of a red light-activated diguanylate cyclase (c-di-GMP synthase) and a blue light-activated c-di-GMP phosphodiesterase (hydrolase) for manipulating intracellular c-di-GMP levels in bacterial cells. We illustrate the application of these enzymes in regulating several c-di-GMP-dependent phenotypes, i.e., motility and biofilm phenotypes in E. coli and chemotactic behavior in the alphaproteobacterium Azospirillum brasilense. We expect these light-activated enzymes to be also useful in regulating c-di-GMP-dependent processes occurring at the fast timescale, for spatial control of bacterial populations, as well as for analyzing c-di-GMP-dependent phenomena at the single-cell level.

Keywords: Biofilm; Chemotaxis; Cyclic dimeric GMP; Diguanylate cyclase; Motility; Optogenetics; Phosphodiesterase; Photoactivation.

MeSH terms

  • Bacteria / enzymology
  • Bacteria / metabolism*
  • Bacteria / radiation effects*
  • Bacterial Physiological Phenomena / radiation effects*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biofilms
  • Biomass
  • Chemotaxis
  • Cyclic GMP / analogs & derivatives*
  • Cyclic GMP / metabolism
  • Enzyme Activators / radiation effects
  • Escherichia coli / physiology
  • Escherichia coli Proteins / metabolism
  • Light*
  • Phosphoric Diester Hydrolases / metabolism
  • Phosphorus-Oxygen Lyases / metabolism
  • Plasmids / genetics
  • Transformation, Bacterial

Substances

  • Bacterial Proteins
  • Enzyme Activators
  • Escherichia coli Proteins
  • bis(3',5')-cyclic diguanylic acid
  • Phosphoric Diester Hydrolases
  • Phosphorus-Oxygen Lyases
  • diguanylate cyclase
  • Cyclic GMP