The Surfeit locus is an unusual cluster of at least 6 housekeeping genes whose organisation is conserved between birds and mammals. We have previously shown that the divergently transcribed Surf-1 and Surf-2 genes are separated by a bi-directional, TATA-less promoter. In mouse, the Surf-1/Surf-2 promoter contains three important factor binding sites: Su1, Su2, and Su3. These sites are conserved between the mouse and human Surf-1/Surf-2 promoters, bind nuclear factors in vitro, and are required for accurate and efficient expression of Surf-1 and Surf-2 in vivo. Using gel retardation assays, methylation interference experiments, and specific antibodies we demonstrate that the Su1 binding factor is the initiator protein YY1. Over-expression of YY1 results in a major stimulation of transcription in the Surf-1 direction and a minor stimulation of transcription in the Surf-2 direction.