Subcellular location characteristics of the Pseudomonas aeruginosa GGDEF protein, WspR, indicate that it produces cyclic-di-GMP in response to growth on surfaces.
The Pseudomonas aeruginosa Wsp signal transduction system produces cyclic-di-GMP (c-di-GMP), an intracellular messenger that stimulates biofilm formation and suppresses motility. The Wsp system is homologous to chemotaxis systems and includes a membrane-bound receptor protein, WspA, and a response regulator GGDEF protein, WspR, that catalyses c-di-GMP synthesis when phosphorylated. We found that the subcellular distributions of fluorescent protein-tagged WspA and WspR differed markedly from their chemotaxis counterparts. WspA-YFP formed patches in cells whereas WspR-YFP was dispersed when unphosphorylated and formed bright cytoplasmic clusters when phosphorylated. WspR formed clusters in cells of a DeltawspF mutant, a genetic background that causes constitutive phosphorylation of WspR, but was dispersed in cells of a wspA mutant, a genetic background necessary for WspR phosphorylation. In addition, WspR mutated at Asp70, its predicted site of phosphorylation, did not form clusters. C-di-GMP synthesis was not required for cluster formation. WspR-YFP was dispersed in liquid-grown wild-type cells, but formed clusters that sometimes appeared and disappeared over the course of a few minutes in cells grown on an agar surface. Our results suggest that the compartmentalized production of c-di-GMP in response to a stimulus associated with growth on a surface is an important functional characteristic of the Wsp system.
Güvener
Zehra Tüzün
ZT
Department of Microbiology, University of Washington, Seattle, WA 98195, USA.
Harwood
Caroline S
CS
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010766:Phosphorylation; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C062025:bis(3',5')-cyclic diguanylic acid; D006152:Cyclic GMP