Yeast promoters URA1 and URA3: Examples of positive control☆

Abstract Transcription of the two unlinked structural genes URA1 and URA3 of Saccharomyces cerevisiae is positively regulated by the gene product PPR 1. We have used S 1 digestion and primer extension mapping to investigate the RNAs produced in different genetic backgrounds: wild-type, ppr 1 deletion mutants, constitutively induced and non-inducible ppr 1 mutants. Results show that each structural gene specifies multiple messenger RNA classes with different 5′-terminal sequences. The basal level of these transcripts does not require a functional PPR 1 gene. Induction of URA1 results from an even increase of the level of synthesis of all the transcripts in contrast to that of URA3 which is effected by selectively increasing the levels of synthesis of one subset of transcripts. The PPR 1-mediated control was also studied in the foreign genetic background of Schizosaccharomyces pombe using autonomously replicating hybrid plasmids carrying the gene URA1 or URA3 along with the regulatory gene PPR 1, either in a constitutive or non-inducible allelic form. The 5′ ends of the transcripts URA1 and URA3 made in S. pombe map upstream from the initiation sites used in S. cerevisiae . In contrast to S. cerevisiae , in S. pombe the URA3 but not URA1 transcripts respond to the PPR 1-induction. We have identified a minimal control region for the PPR 1-specific induction of URA1 , that includes sequences located between the T-A-T-A box and the translation start codon. This region contains sequence features in common with URA3 . There is an extensive alternating Pu:Py region including the T-A-T-A box of both promoters and an eight basepair exact homology; further downstream, there is another 11 base-pair highly conserved sequence which either overlaps or lies in close proximity to the unregulated start sites of URA1 in S. pombe and of URA3 in S. cerevisiae . A positive regulatory model taking into accounts all these observations is presented.