A GAL10-CYC1 Hybrid Yeast Promoter Identifies the GAL4 Regulatory Region As an Upstream Site

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1982 Dec 1

PMID 6760197

Citations 260

Authors

L Guarente

R R Yocum

P Gifford

Affiliations

Soon will be listed here.

Abstract

We have identified the promoter region of the GAL10 gene (whose product is UDP-galactose epimerase) of Saccharomyces cerevisiae; this promoter mediates galactose induction of transcription in conjunction with the product of the GAL4 regulatory gene. This identification was achieved by excising a 365-base-pair fragment of GAL10 leader DNA with a GAL10 proximal endpoint greater than 100 base pairs upstream of the transcriptional start site and substituting it in place of the upstream activation site of the CYC1 (iso-1-cytochrome c) promoter [Guarente, L. & Ptashne, M. (1981) Proc. Natl. Acad. Sci. USA 78, 2199-2203]. The hybrid promoter is composed of DNA encoding CYC1 mRNA start sites and the GAL segment upstream of these sites. This promoter is regulated in a manner analogous to GAL10; i.e., it is induced by galactose and responds to mutations in the GAL4 and GAL80 regulatory loci. The activity of the hybrid promoter requires sequences in the region of the CYC1 mRNA start sites but does not require a precise spacing between these sequences and the GAL segment. The transposed GAL segment appears not to contain sequences that mediate glucose repression. Thus, the picture of the GAL10 promoter that emerges is one of an upstream activation site that responds to the GAL4 product plus galactose, and a region of transcription initiation that may contain sequences that mediate glucose repression. Experiments employing strains inducible (GAL80) or constitutive (gal80) for GAL10 expression indicate that an additional component of glucose repression is inducer exclusion.

Citing Articles

Systematic sequence engineering enhances the induction strength of the glucose-regulated GTH1 promoter of Komagataella phaffii.

Flores-Villegas M, Rebnegger C, Kowarz V, Prielhofer R, Mattanovich D, Gasser B Nucleic Acids Res. 2023; 51(20):11358-11374.

PMID: 37791854 PMC: 10639056. DOI: 10.1093/nar/gkad752.

Natural promoters and promoter engineering strategies for metabolic regulation in Saccharomyces cerevisiae.

He S, Zhang Z, Lu W J Ind Microbiol Biotechnol. 2023; 50(1).

PMID: 36633543 PMC: 9936215. DOI: 10.1093/jimb/kuac029.

Time-resolved microfluidics unravels individual cellular fates during double-strand break repair.

Vertti-Quintero N, Levien E, Poggi L, Amir A, Richard G, Baroud C BMC Biol. 2022; 20(1):269.

PMID: 36464673 PMC: 9720956. DOI: 10.1186/s12915-022-01456-3.

Exploiting the Gal4/UAS System as Plant Orthogonal Molecular Toolbox to Control Reporter Expression in Arabidopsis Protoplasts.

Iacopino S, Licausi F, Giuntoli B Methods Mol Biol. 2022; 2379:99-111.

PMID: 35188658 DOI: 10.1007/978-1-0716-1791-5_6.

Promoter Engineering before and during the Synthetic Biology Era.

Feng X, Marchisio M Biology (Basel). 2021; 10(6).

PMID: 34204069 PMC: 8229000. DOI: 10.3390/biology10060504.

References

Adhya S, Echols H . Glucose effect and the galactose enzymes of Escherichia coli: correlation between glucose inhibition of induction and inducer transport. J Bacteriol. 1966; 92(3):601-8. PMC: 276297. DOI: 10.1128/jb.92.3.601-608.1966. View

DOUGLAS H, Hawthorne D . Regulation of genes controlling synthesis of the galactose pathway enzymes in yeast. Genetics. 1966; 54(3):911-6. PMC: 1211212. DOI: 10.1093/genetics/54.3.911. View

Gascon S, Neumann N, LAMPEN J . Comparative study of the properties of the purified internal and external invertases from yeast. J Biol Chem. 1968; 243(7):1573-7. View

van Wijk R, Ouwehand J, van den Bos T, KONINGSBERGER V . Induction and catabolite repression of alpha-glucosidase synthesis in protoplasts of Saccharomyces carlsbergensis. Biochim Biophys Acta. 1969; 186(1):178-91. DOI: 10.1016/0005-2787(69)90501-2. View

ADAMS B . Induction of galactokinase in Saccharomyces cerevisiae: kinetics of induction and glucose effects. J Bacteriol. 1972; 111(2):308-15. PMC: 251283. DOI: 10.1128/jb.111.2.308-315.1972. View

Nogi Y, Matsumoto K, Toh-E A, Oshima Y . Interaction of super-repressible and dominant constitutive mutations for the synthesis of galactose pathway enzymes in Saccharomyces cerevisiae. Mol Gen Genet. 1977; 152(3):137-44. DOI: 10.1007/BF00268810. View

Saier Jr M, Straud H, Massman L, Judice J, Newman M, Feucht B . Permease-specific mutations in Salmonella typhimurium and Escherichia coli that release the glycerol, maltose, melibiose, and lactose transport systems from regulation by the phosphoenolpyruvate:sugar phosphotransferase system. J Bacteriol. 1978; 133(3):1358-67. PMC: 222173. DOI: 10.1128/jb.133.3.1358-1367.1978. View

Hopper J, Broach J, Rowe L . Regulation of the galactose pathway in Saccharomyces cerevisiae: induction of uridyl transferase mRNA and dependency on GAL4 gene function. Proc Natl Acad Sci U S A. 1978; 75(6):2878-82. PMC: 392668. DOI: 10.1073/pnas.75.6.2878. View

PERLMAN D, Hopper J . Constitutive synthesis of the GAL4 protein, a galactose pathway regulator in Saccharomyces cerevisiae. Cell. 1979; 16(1):89-95. DOI: 10.1016/0092-8674(79)90190-9. View

10.

St John T, Davis R . Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridization. Cell. 1979; 16(2):443-52. DOI: 10.1016/0092-8674(79)90020-5. View

11.

Zitomer R, Montgomery D, Nichols D, Hall B . Transcriptional regulation of the yeast cytochrome c gene. Proc Natl Acad Sci U S A. 1979; 76(8):3627-31. PMC: 383885. DOI: 10.1073/pnas.76.8.3627. View

12.

Matsumoto K, Adachi Y, Toh-E A, Oshima Y . Function of positive regulatory gene gal4 in the synthesis of galactose pathway enzymes in Saccharomyces cerevisiae: evidence that the GAL81 region codes for part of the gal4 protein. J Bacteriol. 1980; 141(2):508-27. PMC: 293654. DOI: 10.1128/jb.141.2.508-527.1980. View

13.

Sherman F, Stewart J, Schweingruber A . Mutants of yeast initiating translation of iso-1-cytochrome c within a region spanning 37 nucleotides. Cell. 1980; 20(1):215-22. DOI: 10.1016/0092-8674(80)90249-4. View

14.

Benoist C, Chambon P . In vivo sequence requirements of the SV40 early promotor region. Nature. 1981; 290(5804):304-10. DOI: 10.1038/290304a0. View

15.

Grosschedl R, Birnstiel M . Spacer DNA sequences upstream of the T-A-T-A-A-A-T-A sequence are essential for promotion of H2A histone gene transcription in vivo. Proc Natl Acad Sci U S A. 1980; 77(12):7102-6. PMC: 350449. DOI: 10.1073/pnas.77.12.7102. View

16.

Gruss P, Dhar R, Khoury G . Simian virus 40 tandem repeated sequences as an element of the early promoter. Proc Natl Acad Sci U S A. 1981; 78(2):943-7. PMC: 319921. DOI: 10.1073/pnas.78.2.943. View

17.

Guarente L, Ptashne M . Fusion of Escherichia coli lacZ to the cytochrome c gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1981; 78(4):2199-203. PMC: 319311. DOI: 10.1073/pnas.78.4.2199. View

18.

Faye G, Leung D, Tatchell K, Hall B, Smith M . Deletion mapping of sequences essential for in vivo transcription of the iso-1-cytochrome c gene. Proc Natl Acad Sci U S A. 1981; 78(4):2258-62. PMC: 319324. DOI: 10.1073/pnas.78.4.2258. View

19.

St John T, Davis R . The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol. 1981; 152(2):285-315. DOI: 10.1016/0022-2836(81)90244-8. View

20.

St John T, Scherer S, McDonell M, Davis R . Deletion analysis of the Saccharomyces GAL gene cluster. Transcription from three promoters. J Mol Biol. 1981; 152(2):317-34. DOI: 10.1016/0022-2836(81)90245-x. View