Figure 3

Novel target genes for Fep1p and Cuf1p. The expression of genes induced during copper and iron starvation and containing one or more putative Cuf1p and Fep1p binding motifs in an 800 base pair promotor region was evaluated by real-time quantitative polymerase chain reaction (qPCR) in strains deleted for either Cuf1p or Fep1p. The fold change in target gene expression in fep1-Δ and cuf1-Δ mutants is shown relative to a wild-type control. The deletion strains were grown in yeast extract (YE) medium, with or without copper or iron chelator added as indicated (± BCS, with or without addition of 100 μmol/l bathocuproinedisulphonate; ± FZ, with or without addition of 300 μmol/l ferrozine). Wild-type control strains were grown in YE medium without metal chelator. Averaged fold changes were obtained by qPCR for two biologic replicates, assayed in duplicate. Significant expression changes (P ≤ 0.05) determined in a two-sided Student's t test are indicated by asterisks. High confidence transcription factor target genes are indicated in red; previously known targets are shown in bold. The maximum observed fold change during the microarray time course, as determined from averaged replicates, is shown in comparison. ^aValue obtained by quantitative real-time PCR.