title: Genetic analysis of the role of yfiR in the ability of Escherichia coli CFT073 to control cellular cyclic dimeric GMP levels and to persist in the urinary tract.
abstract: During urinary tract infections (UTIs), uropathogenic Escherichia coli must maintain a delicate balance between sessility and motility to achieve successful infection of both the bladder and kidneys. Previous studies showed that cyclic dimeric GMP (c-di-GMP) levels aid in the control of the transition between motile and nonmotile states in E. coli. The yfiRNB locus in E. coli CFT073 contains genes for YfiN, a diguanylate cyclase, and its activity regulators, YfiR and YfiB. Deletion of yfiR yielded a mutant that was attenuated in both the bladder and the kidneys when tested in competition with the wild-type strain in the murine model of UTI. A double yfiRN mutant was not attenuated in the mouse model, suggesting that unregulated YfiN activity and likely increased cytoplasmic c-di-GMP levels cause a survival defect. Curli fimbriae and cellulose production were increased in the yfiR mutant. Expression of yhjH, a gene encoding a proven phosphodiesterase, in CFT073 ΔyfiR suppressed the overproduction of curli fimbriae and cellulose and further verified that deletion of yfiR results in c-di-GMP accumulation. Additional deletion of csgD and bcsA, genes necessary for curli fimbriae and cellulose production, respectively, returned colonization levels of the yfiR deletion mutant to wild-type levels. Peroxide sensitivity assays and iron acquisition assays displayed no significant differences between the yfiR mutant and the wild-type strain. These results indicate that dysregulation of c-di-GMP production results in pleiotropic effects that disable E. coli in the urinary tract and implicate the c-di-GMP regulatory system as an important factor in the persistence of uropathogenic E. coli in vivo.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Erica L
authors.initials: EL
authors.lastname: Raterman
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Shapiro
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Stevens
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Schwartz
authors.forename: Rodney A
authors.initials: RA
authors.lastname: Welch
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002482:Cellulose / Q000235:genetics / Q000378:metabolism; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism / Q000382:microbiology; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D004927:Escherichia coli Infections / Q000378:metabolism / Q000382:microbiology*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D010861:Fimbriae, Bacterial / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D006861:Hydrogen Peroxide / Q000378:metabolism; D007501:Iron / Q000378:metabolism; D051379:Mice; D019761:Phosphorus-Oxygen Lyases / Q000235:genetics* / Q000378:metabolism; D014551:Urinary Tract / Q000378:metabolism / Q000382:microbiology*; D014552:Urinary Tract Infections / Q000378:metabolism / Q000382:microbiology*; D014556:Urine / Q000382:microbiology; D056689:Uropathogenic Escherichia coli / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C077481:Crl protein, Bacteria; C062025:bis(3',5')-cyclic diguanylic acid; D002482:Cellulose; D006861:Hydrogen Peroxide; D007501:Iron; D019761:Phosphorus-Oxygen Lyases; C060720:diguanylate cyclase; D006152:Cyclic GMP