title: The concentration of intracellular nickel in Haemophilus influenzae is linked to its surface properties and cell-cell aggregation and biofilm formation.
abstract: Of the known proteins which use nickel as a co-factor, Haemophilus influenzae contains only urease and glyoxalase I (gloA). We have recently reported that this pathogen harbours a unique nickel uptake system (nikKLMQO-nimR). Unusually, the disruption of the nickel uptake system (nikQ or nimR mutants) resulted in cells that aggregated and formed an increased biofilm compared to the wild type cells. Using a gloA mutant strain and urease-specific inhibitor we showed that this phenotype is not due to the loss-of-function of these enzymes. By generating H. influenzae "resting cells" which are enzymatically inactive but maintain their structural integrity we have shown that the cell aggregation in the nikQ/nimR mutants is not due to the loss of enzymatic function. The nikQ mutant was unable to accumulate nickel but the addition of excess nickel did restore intracellular nickel levels and this resulted in the nikQ mutant returning to the wild type "free-living" phenotype; cells with no aggregation and no biofilm formation. We used a range of techniques which showed that the nikQ mutant possesses changes to its cell surface properties. The mutant was more negatively charged than wild type cells as well as being more hydrophobic. Analysis of the outer membrane constituents showed that there were molecular differences. Although the nikQ mutant appears to grow the same as its wild type cell we have shown that there is a change in the "lifestyle" of these nickel limited cells and this induces changes to the surface of the cell to promote cell-cell aggregation and biofilm formation.
authors.affiliation: Research Centre for Infectious Diseases, School of Molecular and Biomedical Sciences, The University of Adelaide, North Terrace Campus, Adelaide, South Australia 5005, Australia.
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Ng
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Kidd
mesh_terms: D001422:Bacterial Adhesion; D018441:Biofilms / Q000254:growth & development*; D003067:Coenzymes / Q000032:analysis*; D004798:Enzymes / Q000235:genetics / Q000378:metabolism; D055786:Gene Knockout Techniques; D006193:Haemophilus influenzae / Q000737:chemistry / Q000502:physiology*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D009532:Nickel / Q000032:analysis*; D013499:Surface Properties
chemical_list: D003067:Coenzymes; D004798:Enzymes; D026901:Membrane Transport Proteins; D009532:Nickel