title: Role of ZnuABC and ZinT in Escherichia coli O157:H7 zinc acquisition and interaction with epithelial cells.
abstract: BACKGROUND
  Zinc is an essential element for all living cells. Recent studies have shown that the ZnuABC zinc uptake system significantly contributes to the ability of several pathogens to multiply in the infected host and cause disease, suggesting that zinc is scarcely available within different tissues of the host. To better understand the role of zinc in bacterial pathogenicity, we have undertaken a functional characterization of the role of the ZnuABC-mediated zinc uptake pathway in enterohemorrhagic Escherichia coli O157:H7.
  
  
  RESULTS
  In this work we have analyzed the expression and the role in metal uptake of ZnuA, the periplasmic component of the ZnuABC transporter, and of ZinT, another periplasmic protein which has been shown to contribute to zinc recruitment. We report that the expression of zinT and znuA, regulated by Zur, is induced in zinc-poor media, and that inactivation of either of the genes significantly decreases E. coli O157:H7 ability to grow in zinc depleted media. We also demonstrate that ZinT and ZnuA have not a redundant function in zinc homeostasis, as the role of ZinT is subordinated to the presence of ZnuA. Moreover, we have found that znuA and zinT are strongly induced in bacteria adhering to cultured epithelial cells and that lack of ZnuA affects the adhesion ability. In addition we have found that a fraction of apo-ZinT can be secreted outside the cell where the protein might sequester environmental zinc, inducing a condition of metal starvation in surrounding cells.
  
  
  CONCLUSIONS
  The here reported results demonstrate that ZnuABC plays a critical role in zinc uptake also in E. coli O157:H7 and that ZinT contributes to the ZnuA-mediated recruitment of zinc in the periplasmic space. Full functionality of the zinc import apparatus is required to facilitate bacterial adhesion to epithelial cells, indicating that the microbial ability to compete with the host cells for zinc binding is critical to establish successful infections. The observation that ZinT can be secreted when it is in the apo-form suggests that its presence in the extracellular environment may somehow contribute to metal uptake or facilitate bacterial colonization of the intestinal epithelia.
authors.affiliation: Biotechnology Service of Service for Biotechnology and Animal Welfare, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. roberta.gabbianelli@iss.it
authors.forename: Roberta
authors.initials: R
authors.lastname: Gabbianelli
authors.forename: Raffaella
authors.initials: R
authors.lastname: Scotti
authors.forename: Serena
authors.initials: S
authors.lastname: Ammendola
authors.forename: Patrizia
authors.initials: P
authors.lastname: Petrarca
authors.forename: Laura
authors.initials: L
authors.lastname: Nicolini
authors.forename: Andrea
authors.initials: A
authors.lastname: Battistoni
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D001422:Bacterial Adhesion; D018938:Caco-2 Cells; D004847:Epithelial Cells / Q000382:microbiology*; D019453:Escherichia coli O157 / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015032:Zinc / Q000378:metabolism*
chemical_list: D018528:ATP-Binding Cassette Transporters; D029968:Escherichia coli Proteins; C470844:ZinT protein, E coli; C488663:ZnuA protein, E coli; C546032:ZnuB protein, E coli; D015032:Zinc