Upstream region required for regulated expression of the glucose-repressible SUC2 gene of Saccharomyces cerevisiae.
Open Access
- 1 December 1984
- journal article
- research article
- Published by Taylor & Francis in Molecular and Cellular Biology
- Vol. 4 (12) , 2750-2757
- https://doi.org/10.1128/mcb.4.12.2750
Abstract
The SUC2 gene produces two mRNAs with different 5' ends that encode two forms of invertase. The 1.9-kilobase mRNA encoding secreted invertase is regulated by glucose repression (carbon catabolite repression), and the 1.8-kilobase mRNA encoding intracellular invertase is produced constitutively at low levels. To identify 5' noncoding sequences essential for regulated expression of SUC2, we constructed in vitro a series of deletions and inserted them into the yeast genome at the chromosomal SUC2 locus. Analysis of the effects of each deletion on SUC2 gene expression identified an upstream region required for derepression of secreted invertase synthesis. The 3' boundary of this region is near -418. The 5' boundary does not appear to be sharply defined, but lies ca. 100 base pairs upstream. A deletion extending from -418 to -140 allowed high-level derepression, indicating that no essential sequences lie between the upstream region and the TATA box at -133 and that the upstream region can be moved 279 base pairs closer to the transcriptional start site. Interactions between the deletions and several unlinked mutations affecting the regulation of SUC2 gene expression were examined. Sequences between -1,900 and -86 are dispensable for expression of the 1.8-kilobase mRNA.This publication has 21 references indexed in Scilit:
- Detection of specific sequences among DNA fragments separated by gel electrophoresisPublished by Elsevier ,2006
- [57] Sequencing end-labeled DNA with base-specific chemical cleavagesPublished by Elsevier ,2004
- [49] Rapid DNA isolations for enzymatic and hybridization analysisPublished by Elsevier ,1980
- Sterile host yeasts (SHY): A eukaryotic system of biological containment for recombinant DNA experimentsGene, 1979
- Replacement of chromosome segments with altered DNA sequences constructed in vitro.Proceedings of the National Academy of Sciences, 1979
- Preparative and analytical purification of DNA from agarose.Proceedings of the National Academy of Sciences, 1979
- Transformation of yeast.Proceedings of the National Academy of Sciences, 1978
- Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase IJournal of Molecular Biology, 1977
- [76] β-d-Fructofuranoside fructohydrolase from yeastPublished by Elsevier ,1975
- Determination of enzymatic activity in polyacrylamide gelsAnalytical Biochemistry, 1969