title: Virulence of a Bordetella pertussis strain expressing a mutant adenylyl cyclase with decreased calmodulin affinity.
abstract: Bordetella pertussis, the pathogen responsible for whooping cough, produces a toxic calmodulin-sensitive adenylyl cyclase which enters animal cells and increases intracellular cAMP. A point mutant of B. pertussis with abolished adenylyl cyclase catalytic activity was over 1000-fold less pathogenic to newborn mice than wild-type bacteria, demonstrating the importance of the adenylyl cyclase for B. pertussis virulence (Gross et al.). The B. pertussis adenylyl cyclase is highly sensitive to calmodulin with an apparent Kd for calmodulin of approximately 1 nM. The importance of this high-affinity calmodulin binding for virulence in vivo was examined by the creation of a B. pertussis point mutant (Trp-242 to Glu-242) with 200-fold lower calmodulin affinity than the native enzyme. This mutant B. pertussis strain retained its virulence in a newborn mouse model of pertussis, but the time course for establishment of a lethal infection in vivo was significantly delayed for the mutant strain. These data illustrate that high-affinity calmodulin binding is not obligatory for the activity of this toxin but is important for the rate for establishment of a lethal infection.
authors.affiliation: Department of Pharmacology, University of Washington School of Medicine, Seattle 98195.
authors.forename: D J
authors.initials: DJ
authors.lastname: Oldenburg
authors.forename: M K
authors.initials: MK
authors.lastname: Gross
authors.forename: A L
authors.initials: AL
authors.lastname: Smith
authors.forename: D R
authors.initials: DR
authors.lastname: Storm
mesh_terms: D000262:Adenylyl Cyclases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001886:Bordetella pertussis / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D002147:Calmodulin / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D017354:Point Mutation; D011485:Protein Binding; D014774:Virulence
chemical_list: D002147:Calmodulin; D000262:Adenylyl Cyclases