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Zinc is an essential trace element required for the function of many enzymes and transcription factors. However, zinc in excess is toxic and intracellular zinc levels must be regulated to maintain homeostasis. In the fission yeast model system Schizosaccharomyces pombe, Loz1 is a transcription factor capable of repressing gene expression under high zinc concentrations. Many Loz1 target genes contain a GNNGATC DNA sequence in their promoter, suggesting that Loz1 directly binds to this Loz1 Response Element (LRE) to repress genetic repression. Using a minimal reporter system, the role of the GNNGATC element in Loz1-dependent regulation has been assessed to determine if this sequence is necessary and sufficient to confer genetic repression under high zinc conditions in-vivo. Additionally, Loz1 target genes pof1 and vel1 were evaluated by lacZ reporter constructs to test whether they were regulated by zinc and Loz1. Both target genes were regulated by zinc in a manner dependent upon Loz1. Mutations in the LRE of the vel1 promoter led to a loss of repression under high zinc concentrations in a wild-type cell indicating that this element is necessary for Loz1 function. In the minimal reporter system, a single LRE element was not sufficient to confer Loz1-mediated repression in the minimal reporter system. However, addition of four LRE were required to repress gene expression in a wild-type cell. This regulation was no longer observed in strains lacking the Loz1 protein. This study shows that Loz1 directly binds to this GNNGATC motif to repress gene expression under high zinc concentrations. Moreover, this study suggests that multiple LRE are required for Loz1 to exert its full effects on zinc-dependent gene expression.