Precise arrangement of factor-binding sites is required for murine CD4 promoter function

The control of CD4 expression is linked to the signaling events that mediate T-cell development and is directly dependent on the CD4 promoter. The CD4 promoter does not contain functionally redundant sites: all four factor-binding sites must be intact to achieve wild-type activity. Here we demonstrate that the precise position of three factor-binding sites relative to each other is essential for promoter activity, indicating that they function together as an inseparable cassette for assembly of the transcription initiation complex. Small changes in either phasing or distance between any two sites in this cassette leads to complete abrogation of promoter function. In addition, we demonstrate that one of the factors that bind the promoter cassette is not present in CD8 SP TC cells. Thus, this factor is a candidate for mediating the relative subclass specificity of CD4 promoter function in activated CD4 SP TH cells.