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Originally published as Genetics Published Articles Ahead of Print on October 16, 2004.

Genetics, Vol. 169, 107-122, January 2005, Copyright © 2005
doi:10.1534/genetics.104.035873

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Genome-Wide Screen for Genes With Effects on Distinct Iron Uptake Activities in Saccharomyces cerevisiae

Emmanuel Lesuisse*,1, Simon A. B. Knight{dagger}, Maïté Courel*, Renata Santos*, Jean-Michel Camadro* and Andrew Dancis{dagger}

* Laboratoire d'Ingéniérie des Protéines et Contrôle Métabolique, Département de Biologie des Génomes, Institut Jacques Monod, Unité Mixte de Recherche 7592 CNRS-Universités Paris 6 and 7, France
{dagger} Department of Medicine, Division of Hematology-Oncology, University of Pennsylvania, Philadelphia, Pennsylvania 19104

1 Corresponding author: Laboratoire d'Ingéniérie des Protéines et Contrôle Métabolique, Institut Jacques Monod, Tour 43, Université Paris 6 and Paris 7, 2 place Jussieu, F-75251 Paris Cedex 05, France.
E-mail: lesuisse{at}ijm.jussieu.fr

We screened a collection of 4847 haploid knockout strains (EUROSCARF collection) of Saccharomyces cerevisiae for iron uptake from the siderophore ferrioxamine B (FOB). A large number of mutants showed altered uptake activities, and a few turned yellow when grown on agar plates with added FOB, indicating increased intracellular accumulation of undissociated siderophores. A subset consisting of 197 knockouts with altered uptake was examined further for regulated activities that mediate cellular uptake of iron from other siderophores or from iron salts. Hierarchical clustering analysis grouped the data according to iron sources and according to mutant categories. In the first analysis, siderophores grouped together with the exception of enterobactin, which grouped with iron salts, suggesting a reductive pathway of iron uptake for this siderophore. Mutant groupings included three categories: (i) high-FOB uptake, high reductase, low-ferrous transport; (ii) isolated high- or low-FOB transport; and (iii) induction of all activities. Mutants with statistically altered uptake activities included genes encoding proteins with predominant localization in the secretory pathway, nucleus, and mitochondria. Measurements of different iron-uptake activities in the yeast knockout collection make possible distinctions between genes with general effects on iron metabolism and those with pathway-specific effects.




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