MUTATIONAL ANALYSIS OF THE ADENOVIRUS E2-EARLY PROMOTER IN FISSION YEAST SCHIZOSACCHAROMYCES-POMBE

The human adenovirus E2-early promoter has-a complex architecture cons isting of overlapping sequences that constitute the major(+1) and mino r(-26) promoters in human cells. In human cells the basal transcriptio n of the major promoter is dependent on 4 cip-acting elements: a TTAAG A motif analogous to the TATA hox, two E2F sites that are present as i nverted repeats, and an ATF/CREB site. It was also demonstrated that t he E2-early promoter was expressed efficiently in the fission yeast Sc hizosaccharomyces pombe and that the major and minor promoters were di fferentially utilized with preferential transcription from the -26 pro moter. In this report the results of an investigation of the E2-early promoter activity in S. pombe, using an additional group of linkerscan mutants that span the E2 promoter, are presented. The efficient expre ssion of the E2-early promoter in yeast was dependent on all 4 cis-act ing elements as monitored by reporter gene expression. However, unlike the situation in human cells, the mutation of the TATA-like element p resent at -50 bps rendered the -26 promoter inactive and was therefore crucial for the maximal promoter function in S. pombe. Ai; in human c ells the wild type. promoter activity was seen in SI pombe when the -8 2 to -92 region was mutated. DNA-protein interaction studies confirmed the presence of ATF and E2F-like transcription factor activities in S . pombe. This report demonstrates the degree of conservation that exis ts between the transcription apparatus of yeast and man.