title: Analysis of a Borrelia burgdorferi phosphodiesterase demonstrates a role for cyclic-di-guanosine monophosphate in motility and virulence.
abstract: The genome of Borrelia burgdorferi encodes a set of genes putatively involved in cyclic-dimeric guanosine monophosphate (cyclic-di-GMP) metabolism. Although BB0419 was shown to be a diguanylate cyclase, the extent to which bb0419 or any of the putative cyclic-di-GMP metabolizing genes impact B. burgdorferi motility and pathogenesis has not yet been reported. Here we identify and characterize a phosphodiesterase (BB0363). BB0363 specifically hydrolyzed cyclic-di-GMP with a K(m) of 0.054 microM, confirming it is a functional cyclic-di-GMP phosphodiesterase. A targeted mutation in bb0363 was constructed using a newly developed promoterless antibiotic cassette that does not affect downstream gene expression. The mutant cells exhibited an altered swimming pattern, indicating a function for cyclic-di-GMP in regulating B. burgdorferi motility. Furthermore, the bb0363 mutant cells were not infectious in mice, demonstrating an important role for cyclic-di-GMP in B. burgdorferi infection. The mutant cells were able to survive within Ixodes scapularis ticks after a blood meal from naïve mice; however, ticks infected with the mutant cells were not able to infect naïve mice. Both motility and infection phenotypes were restored upon genetic complementation. These results reveal an important connection between cyclic-di-GMP, B. burgdorferi motility and Lyme disease pathogenesis. A mechanism by which cyclic-di-GMP influences motility and infection is proposed.
authors.affiliation: Department of Microbiology and Immunology, Brody School of Medicine, 600 Moye Boulevard, Greenville, NC 27834, USA.
authors.forename: Syed Z
authors.initials: SZ
authors.lastname: Sultan
authors.forename: Joshua E
authors.initials: JE
authors.lastname: Pitzer
authors.forename: Michael R
authors.initials: MR
authors.lastname: Miller
authors.forename: Md A
authors.initials: MA
authors.lastname: Motaleb
mesh_terms: D000818:Animals; D025065:Borrelia burgdorferi / Q000201:enzymology* / Q000472:pathogenicity / Q000502:physiology*; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D018884:Ixodes / Q000382:microbiology; D007700:Kinetics; D008124:Locomotion; D008193:Lyme Disease; D051379:Mice; D008809:Mice, Inbred C3H; D050296:Microbial Viability; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000378:metabolism*; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D037521:Virulence Factors; C062025:bis(3',5')-cyclic diguanylic acid; D010727:Phosphoric Diester Hydrolases; D006152:Cyclic GMP