Transcriptional regulator Leu3 of Saccharomyces cerevisiae: Separation of activator and repressor functions

Ji Ying Sze, Eumorphia Remboutsika, Gunter B. Kohlhaw

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

The Leu3 protein of Saccharomyces cerevisiae binds to specific DNA sequences present in the 5′ noncoding region of at least five RNA polymerase II-transcribed genes. Leu3 functions as a transcriptional activator only when the metabolic intermediate α-isopropylmalate is also present. In the absence of α-isopropylmalate, Leu3 causes transcription to be repressed below basal levels. We show here that different portions of the Leu3 protein are responsible for activation and repression. Fusion of the 30 C-terminal residues of Leu3 to the DNA-binding domain of the Ga14 protein created a strong cross-species activator, demonstrating that the short C-terminal region is not only required but also sufficient for transcriptional activation. Using a recently developed Leu3-responsive in vitro transcription assay as a test system for repression (J. Sze, M. Woontner, J. Jaehning, and G. B. Kohlhaw, Science 258:1143-1145, 1992), we show that mutant forms of the Leu3 protein that lack the activation domain still function as repressors. The shortest repressor thus identified had only about 15% of the mass of the full-length Leu3 protein and was centered on the DNA-binding region of Leu3. Implications of this finding for the mechanism of repression are discussed.

Original languageEnglish (US)
Pages (from-to)5702-5709
Number of pages8
JournalMolecular and cellular biology
Volume13
Issue number9
DOIs
StatePublished - Sep 1993
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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