Yeast mRNA initiation sites are determined primarily by specific sequences, not by the distance from the TATA element

We present evidence suggesting that accurate mRNA initiation in yeast cells, unlike their higher eukaryotic counterparts, is determined primarily by specific sequences downstream from the TATA element. First, changing the distance between the his3 TATA element and the initiation region does not affect the sites of initiation or the level of RNA. Second, reciprocal his3-ded1 and ded1-his3 hybrid promoters containing the upstream and TATA elements of one gene fused to the mRNA coding region of the other gene initiate transcription at sites defined by wild-type mRNA coding sequences, not by the distance from the TATA element. Third, when the his3 or ded1 promoter region is fused to position +2 of the his3 gene, transcripts initiated from a position equivalent to +1 are not observed. The results also suggest that the spacing between the TATA element and initiation site is relatively flexible; distance ranging from 40 to 90 bp appear to be functionally acceptable.