Mutations in target DNA elements of yeast HAP1 modulate its transcriptional activity without affecting DNA binding

Ha, Nhuan; Hellauer, Karen; Turcotte, Bernard

doi:10.1093/nar/24.8.1453

Cited by 48 publications

(50 citation statements)

References 42 publications

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“…Dimerization is mediated through a parallel coiled-coil element connected to the zinc cluster by a short extended polypeptide. Recognition can also occur with CGG triplets oriented as direct repeats, as in the case of Hap1p (Zhang and Guarante, 1994;Ha et al, 1996), or as inverted palindromes, as shown recently for Leu3p and Pdr3p . Comparisons of the amino acid sequences of the various C 6 zinc cluster proteins as well as of their DNA targets have appeared recently (Schjerling and Homberg, 1996;Todd and Andrianopoulos, 1997).…”

Section: Introductionmentioning

confidence: 66%

A nonameric core sequence is required upstream of the LYS genes of Saccharomyces cerevisiae for Lys14p‐mediated activation and apparent repression by lysine

Becker

Feller

Alami

et al. 1998

Molecular Microbiology

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SummaryThe expression of the structural genes for lysine (LYS) biosynthesis is controlled by a pathway-specific regulation mediated by the transcriptional activator Lys14 in the presence of ␣-aminoadipate semialdehyde, an intermediate of the pathway acting as a coinducer. Owing to end product inhibition of the first step of the pathway, excess lysine reduces the production of the co-inducer and causes apparent repression of the LYS genes. Analysis of LYS promoters and insertions within an heterologous reporter gene have allowed the characterization of an upstream activating element (UAS LYS ) able to confer Lys14-and ␣-aminoadipate semialdehyde-dependent activation as well as apparent repression by lysine to another yeast gene. This DNA motif is present as one or several copies in the promoters of at least six LYS genes. The consensus sequence derived from the comparison of the UAS LYS showing the highest activation capacities comprises the nonameric core sequence TCCRNYGGA. The RNY sequence of the 3 bp spacer as well as the presence of flanking AT-rich regions on both sides of the core sequence appear essential for optimal activation. Further evidence that this element is the target of Lys14p was provided by the demonstration that Lys14p binds to UAS LYS in vitro. The binding is independent of the presence of the co-inducer and is not affected by lysine. It depends on the integrity of the putative Zn(II) 2 Cys 6 binuclear cluster contained in the Lys14p.

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Section: Introductionmentioning

confidence: 66%

A nonameric core sequence is required upstream of the LYS genes of Saccharomyces cerevisiae for Lys14p‐mediated activation and apparent repression by lysine

Becker

Feller

Alami

et al. 1998

Molecular Microbiology

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“…Most of these factors are vertebrate nuclear receptors, but there is at least another possible example from yeast, the heme-activated Hap1p factor (64). Our study differs from these in that we have defined genetic correlations between structurally distinct Rap1p-DNA complexes and genes involved in various aspects of transcription regulation.…”

Section: Discussionmentioning

confidence: 95%

Alternative Mechanisms of Transcriptional Activation by Rap1p

Idrissi

García-Reyero

Fernández-Larrea

et al. 2001

Journal of Biological Chemistry

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“…One possibility is that GAL4 pc mutants simply have an altered conformation that inhibits the activation domain. Examples of this mechanism comes from work on pc mutants of the glucocorticoid receptor (24) and another Zn 2 Cys 6 protein, HAP1 (25), which indicated that appropriate transcription factor-UAS interactions are necessary for optimal transcriptional activation.…”

Section: Discussionmentioning

confidence: 99%

Alterations in the GAL4 DNA-binding Domain Can Affect Transcriptional Activation Independent of DNA Binding

Corton

Granell

Johnston

1998

Journal of Biological Chemistry

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The GAL4 protein belongs to a large class of fungal transcriptional activator proteins encoding within their DNA-binding domains (DBD) six cysteines that coordinate two atoms of zinc (the Zn 2 Cys 6 domain). In an effort to characterize the interactions between the Zn 2 Cys 6 class transcriptional activator proteins and their DNAbinding sites, we have replaced in the full-length GAL4 protein small regions of the Zn 2 Cys 6 domain with the analogous regions of another Zn 2 Cys 6 protein called PPR1 an activator of pyrimidine biosynthetic genes. Alterations between the first and third cysteines abolished binding to GAL4 (upstream activation sequence of GAL (UAS G )) or PPR1 (upstream acitvation sequence of UAS) DNA-binding sites and severely reduced transcriptional activation in yeast. In contrast, alterations between the third and fourth cysteines had only minor effects on binding to UAS G but led to substantial decreases in activation in both yeast and a mammalian cell line. In the crystal structure of the GAL4 DBD-UAS G complex (Marmorstein, R., Carey, M., Ptashne, M., and Harrison, S. C. (1992) Nature 356, 408 -414), this region is facing away from the DNA, making it likely that there exists within the GAL4 DBD an accessible domain important in activation.The GAL4 protein is an 881-amino acid transcriptional activator of the GAL and MEL1 genes in Saccharomyces cerevisiae. The mechanism by which GAL4 activates transcription is conserved because GAL4 also activates transcription in plant, Drosophila, and mammalian cells if GAL4 DNA-binding sites (UAS G ) 1 are present in the promoter of appropriate reporter genes (reviewed in Ref. 1). GAL4 is a member of a large family of fungal transcriptional activators that contain within their DNA-binding domains a conserved cysteine-rich region encoding six cysteine residues that coordinate two atoms of zinc (the Zn 2 Cys 6 domain). The DNA recognition sequences to which these activators bind have been identified and all have in common a CGG or related triplet in each of the two symmetrically opposed half-sites (2-4). The sequences between the triplets as well as the spacing of the triplet sequence are highly variable, indicating that these variables may in large part determine specific recognition. Based on the extent of homology between the Zn 2 Cys 6 activators, it was hypothesized that the Cys-rich region performs a function common to all members of this class of proteins and the region immediately adjacent, which encodes divergent sequences, determines DNA binding specificity (5). We previously demonstrated that this model is essentially correct (6). All but one (Lys 23 ) of the 28 amino acids in the Zn 2 Cys 6 region of GAL4 can be replaced by the analogous sequences of PPR1, an activator of pyrimidine biosynthetic genes (7), without changing DNA binding specificity. In contrast, replacing the 14 amino acids immediately adjacent to the Zn 2 Cys 6 region resulted in switching the DNA binding specificity to that of PPR1. Similar results were obtained when hybrids of L...

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Mutations in target DNA elements of yeast HAP1 modulate its transcriptional activity without affecting DNA binding

Cited by 48 publications

References 42 publications

A nonameric core sequence is required upstream of the LYS genes of Saccharomyces cerevisiae for Lys14p‐mediated activation and apparent repression by lysine

A nonameric core sequence is required upstream of the LYS genes of Saccharomyces cerevisiae for Lys14p‐mediated activation and apparent repression by lysine

Alternative Mechanisms of Transcriptional Activation by Rap1p

Alterations in the GAL4 DNA-binding Domain Can Affect Transcriptional Activation Independent of DNA Binding

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