Transcriptional regulation of the yeast vacuolar aminopeptidase yscI encoding gene (APE1) by carbon sources

Identification of regulatory proteins that might be involved in carbon catabolite repression of the aminopeptidase I gene of the yeast Saccharomyces cerevisiae

“…c,mtrast to a single upstream activating sequence (UAS) region l~+¢pothetically located between positions -406 and -368, hose removal did not affect significantly fl-galactosidase act~ vity in derepressing conditions, but decreased activity by 1.7-f,,ld in glucose grown cells [2]. To further investigate the existe~lce of such UAS, and taking into account that aminopeptidase ~cI expression is not only under carbon catabolite repression bat also subjected to a strong repression by the nitrogen source c~ the growth medium (Bordallo, J. and Su~,rez-Rendueles, R, v ~anuscript in preparation) we measured fl-galactosidase activi'y in yeast extracts prepared after shifting YPD grown cells to 2 ~ glucose (D) for 4 h. From the results shown in Fig.…”

“…In a previous work we constructed several in-frame fusions of different 5'-truncated versions of the APE1 promoter with the E. coli lacZ gene trying to identify cis-acting regulatory elements controling the transcription of the APE1 gene [2]. c,mtrast to a single upstream activating sequence (UAS) region l~+¢pothetically located between positions -406 and -368, hose removal did not affect significantly fl-galactosidase act~ vity in derepressing conditions, but decreased activity by 1.7-f,,ld in glucose grown cells [2].…”

“…By deletion analysis of APEI-lacZ fusions, a number of promoter regions were shown to be responsible for regulation of APE1 transcription [2]. Based on those experiments, an upstream activating sequence (UAS) was hypothetically located between positions -368 and -406 upstream from the initiating ATG of the gene [2]. A consensus sequence for the binding of the yeast activator protein yAP1 [3] required for normal cadmium tolerance [4] was located within that region.…”

“…Fax: (34) (8) [2] to an A600 nm of 1.00. In growth-shift experiments yeast cells were grown on YPD medium to an A600 nm of 1.00, washed and shifted to 2% glucose (D).…”

“…In growth-shift experiments yeast cells were grown on YPD medium to an A600 nm of 1.00, washed and shifted to 2% glucose (D). The methods used for construction of the APEI-lacZ fusion plasmids, selection of single copy integrants, assay offl-galactosidase and protein determination have been previously described [2].…”