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A systematic analysis reveals an essential role for high-affinity iron uptake system, haemolysin and CFEM domain-containing protein in iron homoeostasis and virulence in Candida glabrata.

The Biochemical journal (2014-07-06)
Vivek Kumar Srivastava, Korivi Jyothiraj Suneetha, Rupinder Kaur
ABSTRACT

Iron is an essential nutrient for all living organisms and human pathogens employ a battery of factors to scavenge iron from the high-affinity iron-binding host proteins. In the present study, we have elucidated, via a candidate gene approach, major iron acquisition and homoeostatic mechanisms operational in an opportunistic human fungal pathogen Candida glabrata. Phenotypic, biochemical and molecular analysis of a set of 13 C. glabrata strains, deleted for proteins potentially implicated in iron metabolism, revealed that the high-affinity reductive iron uptake system is required for utilization of alternate carbon sources and for growth under both in vitro iron-limiting and in vivo conditions. Furthermore, we show for the first time that the cysteine-rich CFEM (common in fungal extracellular membranes) domain-containing cell wall structural protein, CgCcw14, and a putative haemolysin, CgMam3, are essential for maintenance of intracellular iron content, adherence to epithelial cells and virulence. Consistent with their roles in iron homoeostasis, mitochondrial aconitase activity was lower and higher in mutants disrupted for high-affinity iron transport, and haemolysin respectively. Additionally, we present evidence that the mitochondrial frataxin, CgYfh1, is pivotal to iron metabolism. Besides yielding insights into major in vitro and in vivo iron acquisition strategies, our findings establish high-affinity iron uptake mechanisms as critical virulence determinants in C. glabrata.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Iron, ≥99%, reduced, powder (fine)
Sigma-Aldrich
Carbonyl iron, ≥97% Fe basis
Sigma-Aldrich
Iron, foil, thickness 0.25 mm, ≥99.99% trace metals basis
Iron, foil, light tested, 25x25mm, thickness 0.01mm, 99.99+%
Iron, foil, 15mm disks, thickness 0.125mm, as rolled, 99.99+%
Iron, foil, 15mm disks, thickness 0.9mm, as rolled, 99.5%
Iron, foil, 15mm disks, thickness 0.015mm, 99.85%
Iron, foil, 15mm disks, thickness 0.5mm, as rolled, 99.99+%
Iron, foil, 10mm disks, thickness 0.20mm, hard, 99.5%
Iron, foil, 15mm disks, thickness 0.075mm, hard, 99.5%
Iron, foil, 15mm disks, thickness 0.004mm, 99.99+%
Iron, foil, 15mm disks, thickness 0.01mm, 99.85%
Iron, foil, 15mm disks, thickness 0.005mm, 99.85%
Iron, foil, 10mm disks, thickness 0.05mm, as rolled, 99.99+%
Iron, foil, 1m coil, thickness 0.5mm, coil width 49mm, armco« soft ingot 99.8+%
Iron, foil, 25mm disks, thickness 0.9mm, as rolled, 99.5%
Iron, foil, 15mm disks, thickness 1.0mm, as rolled, 99.5%
Iron, foil, 1m coil, thickness 0.125mm, hard, 99.5%
Iron, foil, 15mm disks, thickness 0.25mm, as rolled, 99.99+%
Iron, foil, 15mm disks, thickness 0.007mm, 99.85%
Iron, foil, 10mm disks, thickness 0.1mm, hard, 99.5%
Iron, foil, 1m coil, thickness 0.25mm, hard, 99.5%
Iron, foil, 15mm disks, thickness 0.025mm, as rolled, 99.99+%
Iron, foil, 10mm disks, thickness 0.5mm, as rolled, 99.99+%
Iron, foil, 10mm disks, thickness 0.05mm, hard, 99.5%
Iron, foil, 1m coil, thickness 0.038mm, hard, 99.5%
Iron, foil, 10mm disks, thickness 0.5mm, hard, 99.5%
Iron, foil, 15mm disks, thickness 0.20mm, hard, 99.5%
Iron, foil, 15mm disks, thickness 0.009mm, 99.85%
Iron, foil, 25mm disks, thickness 0.0125mm, 99.85%