Three direct repeats and a TATA-like sequence are required for regulated expression of the human low density lipoprotein receptor gene

The low density lipoprotein receptor is encoded by a 'housekeeping' gene that is transcribed in most mammalian cells and is subject to negative feedback regulation by sterols. To determine the basis for this regulated expression, we performed a transfection analysis with hybrid genes containing up to 6500 base pairs of 5' flanking DNA from the low density lipoprotein receptor gene fused to the coding region of the bacterial chloramphenicol acetyltransferase gene. These studies identified a 177-base pair fragment of 5' flanking DNA that is sufficient for expression as well as negative regulation by sterols. The positive elements within this region were further defined by analysis of a series of 15 mutations in which overlapping 10-base pair segments were scrambled by site-specific mutagenesis. These studies identified the positive elements as three imperfect direct repeats of 16 base pairs and a TATA-like sequence. The three repeats contain a sequence that is homologous to the consensus DNA sequence recognized by transcription factor Sp1.