Identification of an Upstream Activating Sequence and an Upstream Repressible Sequence of the Pyruvate Kinase Gene of the Yeast Saccharomyces cerevisiae

Abstract

To clarify carbon source-dependent control of the glycolytic pathway in the yeast Saccharomyces cerevisiae, we have initiated a study of transcriptional regulation of the pyruvate kinase gene (PYK). By deletion analysis of the 5′-noncoding region of the PYK gene, we have identified an upstream activating sequence (UAS_PYK1) located between 634 and 653 nucleotides upstream of the initiating ATG codon. The promoter activity of the PYK 5′-noncoding region was abolished when the sequence containing the UAS_PYK1 was deleted from the region. Synthetic UAS_PYK1 (26mer), in either orientation, was able to restore the transcriptional activity of UAS-depleted mutants when placed upstream of the TATA sequence located at −199 (ATG as +1). While the UAS_PYK1 was required for basal to intermediate levels of transcriptional activation, a sequence between −714 and −811 was found to be necessary for full activation. On the other hand, a sequence between −344 and −468 was found to be responsible for transcriptional repression of the PYK gene when yeast cells were grown on nonfermentable carbon sources. This upstream repressible sequence also repressed transcription, although to a lesser extent, when glucose was present in the medium. The possible mechanism for carbon source-dependent regulation of PYK expression through these cis-acting regulatory elements is discussed.