Binding and activation by the zinc cluster transcription factors of Saccharomyces cerevisiae

Redefining the UASGABA and its interaction with Uga3p

Anu M. Idicula, Gregory L. Blatch, Terrance Cooper, Rosemary A. Dorrington

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Uga3p, a member of zinc binuclear cluster transcription factor family, is required for γ-aminobutyric acid-dependent transcription of the UGA genes in Saccharomyces cerevisiae. Members of this family bind to CGG triplets with the spacer region between the triplets being an important specificity determinant. A conserved 19-nucleotide activation element in certain UGA gene promoter regions contains a CCGN4CGG-everted repeat proposed to be the binding site of Uga3p, UASGABA. The function of conserved nucleotides flanking the everted repeat has not been rigorously investigated. The interaction of Uga3p with UASGABA was characterized in terms of binding in vitro and transcriptional activation of lacZ reporter genes in vivo. Electromobility shift assays using mutant UASGABA sequences and heterologously produced full-length Uga3p demonstrated that UASGABA consists of two independent Uga3p binding sites. Simultaneous occupation of both Uga3p binding sites of UASGABA with high affinity is essential for GABA-dependent transcriptional activation in vivo. We present evidence that the two Uga3p molecules bound to UASGABA probably interact with each other and show that Uga3p(1-124), previously used for binding studies, is not functionally equivalent to the full-length protein with respect to binding in vitro. We propose that the Uga3p binding site is an asymmetric site of 5′-SGCGGN-WTTT-3′ (S = G or C, W = A, or T and n = no nucleotide or G). However, UASGABA, is a palindrome containing two asymmetric Uga3p binding sites.

Original languageEnglish (US)
Pages (from-to)45977-45983
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number48
DOIs
StatePublished - Nov 29 2002

Fingerprint

Yeast
Saccharomyces cerevisiae
Zinc
Transcription Factors
Chemical activation
Binding Sites
Nucleotides
Genes
Transcriptional Activation
Aminobutyrates
Lac Operon
Transcription
Reporter Genes
Occupations
Genetic Promoter Regions
gamma-Aminobutyric Acid
Assays
Molecules
Proteins
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Binding and activation by the zinc cluster transcription factors of Saccharomyces cerevisiae : Redefining the UASGABA and its interaction with Uga3p. / Idicula, Anu M.; Blatch, Gregory L.; Cooper, Terrance; Dorrington, Rosemary A.

In: Journal of Biological Chemistry, Vol. 277, No. 48, 29.11.2002, p. 45977-45983.

Research output: Contribution to journalArticle

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