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Using quantitative S1 nuclease analysis and recombinants which contain the SV40 early promoter region linked to the rabbit beta-globin gene coding sequence, we have studied the effect of deletion, inversion and point mutations, located within the 21 bp repeat region, on initiation of transcription from the early-early and late-early startsites in the absence of T-antigen. Our data establish unequivocally that the six GC-rich repeats present in the 21 bp repeat region are essential elements of both the early-early and late-early promoters and that they are not redundant, since mutations, which affect only the GC-rich repeat most proximal to the TATA box, decrease drastically the activity of the early-early promoter, but increase that of the late-early promoter. On the other hand, the four GC-rich repeats most proximal to the 72 bp repeat are common elements of the two overlapping early-early and late-early promoters. Our results, which confirm that the early-early promoter is stronger than the late-early one, also support our previous suggestion that they are in competition for the transcriptional machinery. The general organization of the SV40 early promoter region is discussed.

nucleic acids research

Mutational dissection of the 21 bp repeat region of the SV40 early promoter reveals that it contains overlapping elements of the early-early and late-early promoters