Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1993 Sep 11;21(18):4378–4382. doi: 10.1093/nar/21.18.4378

Autoregulation of GAL4 transcription is essential for rapid growth of Kluyveromyces lactis on lactose and galactose.

M Czyz 1, M M Nagiec 1, R C Dickson 1
PMCID: PMC310076  PMID: 8414996

Abstract

Transcriptional induction of genes in the lactose-galactose regulon of the yeast Kluyveromyces lactis requires the GAL4 transcription activator protein. Previous data indicated that the concentration of GAL4 was tightly regulated under basal, inducing, and glucose repressing conditions but the mechanisms were unknown. In this paper we demonstrate that transcription of the GAL4 gene (KI-GAL4) increases 3- to 4-fold during induction of the regulon. This increase requires a KI-GAL4 binding site, UASG, in front of the KI-GAL4 gene, indicating that the KI-GAL4 protein autoregulates transcription of its own gene. Our data demonstrate that the autoregulatory circuit is essential for full induction of the lactose-galactose regulon and, hence, for rapid growth on lactose or galactose. Other data indicate that basal transcription of the KI-GAL4 gene is governed by unidentified promoter elements. The existence of the autoregulatory circuit reveals an important difference between the lactose-galactose regulon and its homologue in Saccharomyces cerevisiae, the melibiose-galactose regulon. This difference may have evolved in response to different selective pressures encountered by the two organisms.

Full text

PDF
4378

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alton N. K., Vapnek D. Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9. Nature. 1979 Dec 20;282(5741):864–869. doi: 10.1038/282864a0. [DOI] [PubMed] [Google Scholar]
  2. Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
  3. Breunig K. D. Glucose repression of LAC gene expression in yeast is mediated by the transcriptional activator LAC9. Mol Gen Genet. 1989 Apr;216(2-3):422–427. doi: 10.1007/BF00334386. [DOI] [PubMed] [Google Scholar]
  4. Breunig K. D., Kuger P. Functional homology between the yeast regulatory proteins GAL4 and LAC9: LAC9-mediated transcriptional activation in Kluyveromyces lactis involves protein binding to a regulatory sequence homologous to the GAL4 protein-binding site. Mol Cell Biol. 1987 Dec;7(12):4400–4406. doi: 10.1128/mcb.7.12.4400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dickson R. C., Barr K. Characterization of lactose transport in Kluyveromyces lactis. J Bacteriol. 1983 Jun;154(3):1245–1251. doi: 10.1128/jb.154.3.1245-1251.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dickson R. C., Sheetz R. M., Lacy L. R. Genetic regulation: yeast mutants constitutive for beta-galactosidase activity have an increased level of beta-galactosidase messenger ribonucleic acid. Mol Cell Biol. 1981 Nov;1(11):1048–1056. doi: 10.1128/mcb.1.11.1048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Falvey E., Schibler U. How are the regulators regulated? FASEB J. 1991 Mar 1;5(3):309–314. doi: 10.1096/fasebj.5.3.2001790. [DOI] [PubMed] [Google Scholar]
  8. Griggs D. W., Johnston M. Regulated expression of the GAL4 activator gene in yeast provides a sensitive genetic switch for glucose repression. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8597–8601. doi: 10.1073/pnas.88.19.8597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Guarente L. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 1983;101:181–191. doi: 10.1016/0076-6879(83)01013-7. [DOI] [PubMed] [Google Scholar]
  10. Gödecke A., Zachariae W., Arvanitidis A., Breunig K. D. Coregulation of the Kluyveromyces lactis lactose permease and beta-galactosidase genes is achieved by interaction of multiple LAC9 binding sites in a 2.6 kbp divergent promoter. Nucleic Acids Res. 1991 Oct 11;19(19):5351–5358. doi: 10.1093/nar/19.19.5351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Halvorsen Y. D., Nandabalan K., Dickson R. C. Identification of base and backbone contacts used for DNA sequence recognition and high-affinity binding by LAC9, a transcription activator containing a C6 zinc finger. Mol Cell Biol. 1991 Apr;11(4):1777–1784. doi: 10.1128/mcb.11.4.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Heus J. J., Zonneveld B. J., Steensma H. Y., Van den Berg J. A. Centromeric DNA of Kluyveromyces lactis. Curr Genet. 1990 Dec;18(6):517–522. doi: 10.1007/BF00327022. [DOI] [PubMed] [Google Scholar]
  13. Karin M. Signal transduction from cell surface to nucleus in development and disease. FASEB J. 1992 May;6(8):2581–2590. doi: 10.1096/fasebj.6.8.1317309. [DOI] [PubMed] [Google Scholar]
  14. Kuger P., Gödecke A., Breunig K. D. A mutation in the Zn-finger of the GAL4 homolog LAC9 results in glucose repression of its target genes. Nucleic Acids Res. 1990 Feb 25;18(4):745–751. doi: 10.1093/nar/18.4.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kuzhandaivelu N., Jones W. K., Martin A. K., Dickson R. C. The signal for glucose repression of the lactose-galactose regulon is amplified through subtle modulation of transcription of the Kluyveromyces lactis Kl-GAL4 activator gene. Mol Cell Biol. 1992 May;12(5):1924–1931. doi: 10.1128/mcb.12.5.1924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Leonardo J. M., Bhairi S. M., Dickson R. C. Identification of upstream activator sequences that regulate induction of the beta-galactosidase gene in Kluyveromyces lactis. Mol Cell Biol. 1987 Dec;7(12):4369–4376. doi: 10.1128/mcb.7.12.4369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Meyer J., Walker-Jonah A., Hollenberg C. P. Galactokinase encoded by GAL1 is a bifunctional protein required for induction of the GAL genes in Kluyveromyces lactis and is able to suppress the gal3 phenotype in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Nov;11(11):5454–5461. doi: 10.1128/mcb.11.11.5454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mylin L. M., Gerardot C. J., Hopper J. E., Dickson R. C. Sequence conservation in the Saccharomyces and Kluveromyces GAL11 transcription activators suggests functional domains. Nucleic Acids Res. 1991 Oct 11;19(19):5345–5350. doi: 10.1093/nar/19.19.5345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mylin L. M., Johnston M., Hopper J. E. Phosphorylated forms of GAL4 are correlated with ability to activate transcription. Mol Cell Biol. 1990 Sep;10(9):4623–4629. doi: 10.1128/mcb.10.9.4623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Parthun M. R., Jaehning J. A. A transcriptionally active form of GAL4 is phosphorylated and associated with GAL80. Mol Cell Biol. 1992 Nov;12(11):4981–4987. doi: 10.1128/mcb.12.11.4981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ruzzi M., Breunig K. D., Ficca A. G., Hollenberg C. P. Positive regulation of the beta-galactosidase gene from Kluyveromyces lactis is mediated by an upstream activation site that shows homology to the GAL upstream activation site of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Mar;7(3):991–997. doi: 10.1128/mcb.7.3.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sadowski I., Niedbala D., Wood K., Ptashne M. GAL4 is phosphorylated as a consequence of transcriptional activation. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10510–10514. doi: 10.1073/pnas.88.23.10510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Seed B., Sheen J. Y. A simple phase-extraction assay for chloramphenicol acyltransferase activity. Gene. 1988 Jul 30;67(2):271–277. doi: 10.1016/0378-1119(88)90403-9. [DOI] [PubMed] [Google Scholar]
  24. Wray L. V., Jr, Witte M. M., Dickson R. C., Riley M. I. Characterization of a positive regulatory gene, LAC9, that controls induction of the lactose-galactose regulon of Kluyveromyces lactis: structural and functional relationships to GAL4 of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Mar;7(3):1111–1121. doi: 10.1128/mcb.7.3.1111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zachariae W., Breunig K. D. Expression of the transcriptional activator LAC9 (KlGAL4) in Kluyveromyces lactis is controlled by autoregulation. Mol Cell Biol. 1993 May;13(5):3058–3066. doi: 10.1128/mcb.13.5.3058. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES