Modulation of APRT transcription by altering spacing between cis-regulatory elements

Three regulatory regions on the promoter of CHO adenine phosphoribosyltransferase gene (APRT) were identified. Spacing constraints between these regions were analyzed. With normal spacing, region II (-33 to +19), which separates regions I (-101 to -53) and III (+56 to +85), is critical for APRT transcription. However, when regions I and III are artificially placed in proximity to each other (region II deleted), they are able to drive transcription as efficiently as the wild-type APRT promoter. Neither region I nor III alone is sufficient for efficient transcription. As the spacing between the two regions increases, the transcription decreases. Region I may activate transcription in two ways: through a stringent sequence-specific manner (as in the transcription mediated by regions I and III) and through a manner with relaxed requirement for sequence specificity (as in the transcription from the wild-type APRT promoter).