Home LiteratureArticle Details
PMID: 3540610 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Efficient expression of the Saccharomyces cerevisiae PGK gene depends on an upstream activation sequence but does not require TATA sequences.

Molecular and cellular biology ·Vol. 6 ·No. 12 ·1986-12-00 ·Pages 4335-43

Ogden JE, Stanway C, Kim S, Mellor J, Kingsman AJ, Kingsman SM

Abstract

The Saccharomyces cerevisiae PGK (phosphoglycerate kinase) gene encodes one of the most abundant mRNA and protein species in the cell. To identify the promoter sequences required for the efficient expression of PGK, we undertook a detailed internal deletion analysis of the 5' noncoding region of the gene. Our analysis revealed that PGK has an upstream activation sequence (UASPGK) located between 402 and 479 nucleotides upstream from the initiating ATG sequence which is required for full transcriptional activity. Deletion of this sequence caused a marked reduction in the levels of PGK transcription. We showed that PGK has no requirement for TATA sequences; deletion of one or both potential TATA sequences had no effect on either the levels of PGK expression or the accuracy of transcription initiation. We also showed that the UASPGK functions as efficiently when in the inverted orientation and that it can enhance transcription when placed upstream of a TRP1-IFN fusion gene comprising the promoter of TRP1 fused to the coding region of human interferon alpha-2.

MeSH Terms
Base Sequence Chromosome Deletion Genes Genes, Fungal Genes, Regulator Mutation Phosphoglycerate Kinase/genetics Plasmids Promoter Regions, Genetic RNA, Messenger/genetics Saccharomyces cerevisiae/enzymology,genetics Transcription, Genetic
Chemicals
RNA, Messenger Phosphoglycerate Kinase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ogden J E
Stanway C
Kim S
Mellor J
Kingsman A J
Kingsman S M
References (54)
54 references, click to expand
  1. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Aug;4(8):1440-8 PMID: 6092912
  2. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
    J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
  3. Efficient synthesis of enzymatically active calf chymosin in Saccharomyces cerevisiae.
    Gene. 1983 Sep;24(1):1-14 PMID: 6313478
  4. The ovalbumin gene-sequence of putative control regions.
    Nucleic Acids Res. 1980 Jan 11;8(1):127-42 PMID: 6243777
  5. Expression of the human interferon-gamma cDNA in yeast.
    Nucleic Acids Res. 1983 Mar 25;11(6):1819-37 PMID: 6300780
  6. DNA sequences necessary for transcription of the rabbit beta-globin gene in vivo.
    Nature. 1982 Jan 14;295(5845):120-6 PMID: 6276753
  7. A repressor (MAT alpha 2 Product) and its operator control expression of a set of cell type specific genes in yeast.
    Cell. 1985 Aug;42(1):237-47 PMID: 3893743
  8. Specific protein binding to far upstream activating sequences in polymerase II promoters.
    Proc Natl Acad Sci U S A. 1985 Jan;82(1):43-7 PMID: 3881758
  9. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201-5 PMID: 6159641
  10. Upstream region required for regulated expression of the glucose-repressible SUC2 gene of Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Dec;4(12):2750-7 PMID: 6396505
  11. Enhancer elements.
    Cell. 1983 Jun;33(2):313-4 PMID: 6305503
  12. The yeast his3 promoter contains at least two distinct elements.
    Proc Natl Acad Sci U S A. 1982 Dec;79(23):7385-9 PMID: 6760196
  13. A positive regulatory site and a negative regulatory site control the expression of the Saccharomyces cerevisiae CYC7 gene.
    Mol Cell Biol. 1984 Oct;4(10):2023-30 PMID: 6095036
  14. Overlap hybridization screening: isolation and characterization of overlapping DNA fragments surrounding the leu2 gene on yeast chromosome III.
    Gene. 1979 Feb;5(2):111-26 PMID: 376402
  15. A region flanking the GAL7 gene and a binding site for GAL4 protein as upstream activating sequences in yeast.
    J Mol Biol. 1985 Dec 20;186(4):821-4 PMID: 3912516
  16. Genetic properties and chromatin structure of the yeast gal regulatory element: an enhancer-like sequence.
    Proc Natl Acad Sci U S A. 1984 Dec;81(24):7865-9 PMID: 6096864
  17. Negative control at a distance mediates catabolite repression in yeast.
    Nature. 1985 Oct 31-Nov 6;317(6040):822-4 PMID: 3903516
  18. Upstream region of the SUC2 gene confers regulated expression to a heterologous gene in Saccharomyces cerevisiae.
    Mol Cell Biol. 1985 Oct;5(10):2521-6 PMID: 3939253
  19. Isolation and identification of yeast messenger ribonucleic acids coding for enolase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase.
    Biochemistry. 1978 Nov 14;17(23):4900-7 PMID: 363146
  20. Each of three "TATA elements" specifies a subset of the transcription initiation sites at the CYC-1 promoter of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1985 Dec;82(24):8562-6 PMID: 3001709
  21. Yeast promoters: positive and negative elements.
    Cell. 1984 Apr;36(4):799-800 PMID: 6368004
  22. A rapid boiling method for the preparation of bacterial plasmids.
    Anal Biochem. 1981 Jun;114(1):193-7 PMID: 6269464
  23. The primary structure of the Saccharomyces cerevisiae gene for 3-phosphoglycerate kinase.
    Nucleic Acids Res. 1982 Dec 11;10(23):7791-808 PMID: 6296791
  24. Isolation of yeast DNA.
    Methods Cell Biol. 1975;12:39-44 PMID: 1105072
  25. Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae.
    Nucleic Acids Res. 1982 Apr 24;10(8):2625-37 PMID: 6281737
  26. The relationship between the "TATA" sequence and transcription initiation sites at the HIS4 gene of Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1985 Dec;82(24):8557-61 PMID: 3909147
  27. The identification and high level expression of a protein encoded by the yeast Ty element.
    EMBO J. 1984 May;3(5):1115-9 PMID: 6329736
  28. A method for the recovery of DNA from agarose gels.
    Nucleic Acids Res. 1978 Jul;5(7):2321-32 PMID: 673856
  29. Sequence of the promoter region of the gene for human X-linked 3-phosphoglycerate kinase.
    Gene. 1984 Dec;32(3):409-17 PMID: 6099325
  30. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  31. The synthesis and in vivo assembly of functional antibodies in yeast.
    Nature. 1985 Apr 4-10;314(6010):446-9 PMID: 3920532
  32. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
    Nucleic Acids Res. 1984 Sep 25;12(18):7035-56 PMID: 6091052
  33. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  34. GCN4 protein, synthesized in vitro, binds HIS3 regulatory sequences: implications for general control of amino acid biosynthetic genes in yeast.
    Cell. 1985 Nov;43(1):177-88 PMID: 3907851
  35. Saccharomyces cerevisiae CYC1 mRNA 5'-end positioning: analysis by in vitro mutagenesis, using synthetic duplexes with random mismatch base pairs.
    Mol Cell Biol. 1985 Dec;5(12):3545-51 PMID: 3915780
  36. Chromosome Mapping in Saccharomyces: Centromere-Linked Genes.
    Genetics. 1960 Aug;45(8):1085-110 PMID: 17247984
  37. Glycolysis mutants in Saccharomyces cerevisiae.
    Genetics. 1978 Jan;88(1):1-11 PMID: 147195
  38. Transformation of yeast.
    Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929-33 PMID: 347451
  39. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  40. A synthetic HIS4 regulatory element confers general amino acid control on the cytochrome c gene (CYC1) of yeast.
    Proc Natl Acad Sci U S A. 1985 Jan;82(2):498-502 PMID: 2982161
  41. Saccharomyces cerevisiae GAL1-GAL10 divergent promoter region: location and function of the upstream activating sequence UASG.
    Mol Cell Biol. 1984 Nov;4(11):2467-78 PMID: 6392852
  42. Regulated high efficiency expression of human interferon-alpha in Saccharomyces cerevisiae.
    EMBO J. 1982;1(5):603-8 PMID: 6329694
  43. Multiple control elements in the TRP1 promoter of Saccharomyces cerevisiae.
    Mol Cell Biol. 1986 Dec;6(12):4251-8 PMID: 3025648
  44. Factors affecting heterologous gene expression in Saccharomyces cerevisiae.
    Gene. 1985;33(2):215-26 PMID: 3888784
  45. Heme regulates transcription of the CYC1 gene of S. cerevisiae via an upstream activation site.
    Cell. 1983 Apr;32(4):1279-86 PMID: 6301690
  46. Upstream activation sites of the CYC1 gene of Saccharomyces cerevisiae are active when inverted but not when placed downstream of the "TATA box".
    Proc Natl Acad Sci U S A. 1984 Dec;81(24):7860-4 PMID: 6096863
  47. Point mutations implicate repeated sequences as essential elements of the CYC7 negative upstream site in Saccharomyces cerevisiae.
    Mol Cell Biol. 1985 Nov;5(11):2951-8 PMID: 3018489
  48. Characterization of two types of yeast ribosomal DNA genes.
    J Bacteriol. 1978 Apr;134(1):295-305 PMID: 348684
  49. Transcription and regulatory signals at the mating type locus in yeast.
    Cell. 1984 Jul;37(3):969-78 PMID: 6378388
  50. Specific DNA binding of GAL4, a positive regulatory protein of yeast.
    Cell. 1985 Apr;40(4):767-74 PMID: 3886158
  51. Yeast mRNA initiation sites are determined primarily by specific sequences, not by the distance from the TATA element.
    EMBO J. 1985 Dec 1;4(12):3273-80 PMID: 3912167
  52. Use of lacZ fusions to delimit regulatory elements of the inducible divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Oct;4(10):1985-98 PMID: 6390181
  53. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene.
    Cell. 1983 Oct;34(3):865-79 PMID: 6313211
  54. Organization and expression of eucaryotic split genes coding for proteins.
    Annu Rev Biochem. 1981;50:349-83 PMID: 6791577
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.6.12.4335-4343.1986
Published
1986-12-00
Pages
4335-43
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367215
Subset
IM
Databases
GENBANK
M14679, X05969
External Links
PubMed source DOI: 10.1128/mcb.6.12.4335-4343.1986 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2