title: Dot/Icm type IVB secretion system requirements for Coxiella burnetii growth in human macrophages.
abstract: Central to Q fever pathogenesis is replication of the causative agent, Coxiella burnetii, within a phagolysosome-like parasitophorous vacuole (PV) in mononuclear phagocytes. C. burnetii modulates PV biogenesis and other host cell functions, such as apoptotic signaling, presumably via the activity of proteins delivered to the host cytosol by a Dot/Icm type IVB secretion system (T4BSS). In this study, we utilized a C. burnetii strain carrying IcmD inactivated by the Himar1 transposon to investigate the requirements for Dot/Icm function in C. burnetii parasitism of human THP-1 macrophage-like cells. The icmD::Tn mutant failed to secrete characterized T4BSS substrates, a defect that correlated with deficient replication, PV development, and apoptosis protection. Restoration of type IVB secretion and intracellular growth of the icmD::Tn mutant required complementation with icmD, -J, and -B, indicating a polar effect of the transposon insertion on downstream dot/icm genes. Induction of icmDJB expression at 1 day postinfection resulted in C. burnetii replication and PV generation. Collectively, these data prove that T4BSS function is required for productive infection of human macrophages by C. burnetii. However, illustrating the metabolic flexibility of C. burnetti, the icmD::Tn mutant could replicate intracellularly when sequestered in a PV generated by wild-type bacteria, where Dot/Icm function is provided in trans, and within a phenotypically similar PV generated by the protozoan parasite Leishmania amazonensis, where host cells are devoid of Dot/Icm T4BSS effector proteins.
authors.affiliation: Coxiella Pathogenesis Section, Laboratory of Intracellular Parasites, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Beare
authors.forename: Stacey D
authors.initials: SD
authors.lastname: Gilk
authors.forename: Charles L
authors.initials: CL
authors.lastname: Larson
authors.forename: Joshua
authors.initials: J
authors.lastname: Hill
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Stead
authors.forename: Anders
authors.initials: A
authors.lastname: Omsland
authors.forename: Diane C
authors.initials: DC
authors.lastname: Cockrell
authors.forename: Dale
authors.initials: D
authors.lastname: Howe
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Voth
authors.forename: Robert A
authors.initials: RA
authors.lastname: Heinzen
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D058947:Bacterial Secretion Systems; D002460:Cell Line; D016997:Coxiella burnetii / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology*; D021381:Protein Transport; D011778:Q Fever / Q000378:metabolism / Q000382:microbiology*; D014617:Vacuoles / Q000378:metabolism / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems