title: Coordinated zinc homeostasis is essential for the wild-type virulence of Brucella abortus.
abstract: Metal homeostasis in bacterial cells is a highly regulated process requiring intricately coordinated import and export, as well as precise sensing of intracellular metal concentrations. The uptake of zinc (Zn) has been linked to the virulence of Brucella abortus; however, the capacity of Brucella strains to sense Zn levels and subsequently coordinate Zn homeostasis has not been described. Here, we show that expression of the genes encoding the zinc uptake system ZnuABC is negatively regulated by the Zn-sensing Fur family transcriptional regulator, Zur, by direct interactions between Zur and the promoter region of znuABC. Moreover, the MerR-type regulator, ZntR, controls the expression of the gene encoding the Zn exporter ZntA by binding directly to its promoter. Deletion of zur or zntR alone did not result in increased zinc toxicity in the corresponding mutants; however, deletion of zntA led to increased sensitivity to Zn but not to other metals, such as Cu and Ni, suggesting that ZntA is a Zn-specific exporter. Strikingly, deletion of zntR resulted in significant attenuation of B. abortus in a mouse model of chronic infection, and subsequent experiments revealed that overexpression of zntA in the zntR mutant is the molecular basis for its decreased virulence.
  
  
  OBJECTIVE
  The importance of zinc uptake for Brucella pathogenesis has been demonstrated previously, but to date, there has been no description of how overall zinc homeostasis is maintained and genetically controlled in the brucellae. The present work defines the predominant zinc export system, as well as the key genetic regulators of both zinc uptake and export in Brucella abortus. Moreover, the data show the importance of precise coordination of the zinc homeostasis systems as disregulation of some elements of these systems leads to the attenuation of Brucella virulence in a mouse model. Overall, this study advances our understanding of the essential role of zinc in the pathogenesis of intracellular bacteria.
authors.affiliation: Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: Lauren M
authors.initials: LM
authors.lastname: Sheehan
authors.affiliation: Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia, USA.
authors.forename: James A
authors.initials: JA
authors.lastname: Budnick
authors.affiliation: Department of Microbiology and Immunology, East Carolina University, Brody School of Medicine, Greenville, North Carolina, USA.
authors.forename: R Martin
authors.initials: RM
authors.lastname: Roop
authors.affiliation: Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia, USA caswellc@vt.edu.
authors.forename: Clayton C
authors.initials: CC
authors.lastname: Caswell
mesh_terms: D000818:Animals; D002003:Brucella abortus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D002006:Brucellosis / Q000382:microbiology / Q000473:pathology; D004195:Disease Models, Animal; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D006706:Homeostasis; D051379:Mice; D014157:Transcription Factors / Q000378:metabolism; D014774:Virulence; D015032:Zinc / Q000378:metabolism* / Q000633:toxicity
chemical_list: D014157:Transcription Factors; D015032:Zinc