New strategies to identify genes regulated by Zap1. A) Identifying genes affected by moderate zinc deficiency. Experiments E1, E2, and E3 were combined to identify genes that showed increased expression in zinc-limited cells (E1), wild type cells vs. zap1Δ mutant cells in low zinc (E2), and Zap1TC-expressing cells in high zinc (E3). B) The ZRE sequences in the promoters of the previously identified 46 Zap1 target genes  were aligned and a logo was built using WebLogo. C) Experiments E3 and E4 were combined to identify Zap1 targets that respond to severe zinc deficiency. Regulatory Sequence Analysis Tools (RSAT) was used to identify potential ZREs in the promoters of co-regulated genes.