Yeast Transcriptional Regulator Leu3p

Wang, Dake; Feng, Zheng; Holmberg, Steen; Kohlhaw, Gunter B.

doi:10.1074/jbc.274.27.19017

Cited by 42 publications

(14 citation statements)

References 33 publications

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“…Leu3p function has been investigated thoroughly by Kohlhaw and co-workers (Wang et al 1999 and references therein). Leu3p binding is required to obtain the full BAP2 promoter activity seen in SC medium (Table 3).…”

Section: Discussionmentioning

confidence: 98%

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Transcriptional regulation of the Saccharomyces cerevisiae amino acid permease gene BAP2

et al. 2001

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Uptake of branched-chain amino acids by Saccharomyces cerevisiae from media containing a preferred nitrogen source is mediated by the permeases encoded by BAP2, BAP3, and VAP1/TAT1. The transcriptional activity of the BAP2 promoter is affected by a number of genes, including SSY1, which encodes an amino acid permease homologue that is necessary for transcription of BAP2. Other genes that control BAP2 encode known (Leu3p, Tup1p) and putative (Stp1p, Stp2p) transcription factors. We present evidence that the zinc-finger proteins Stp1p and Stp2p bind directly to the BAP2 promoter. Binding of Stplp to the BAP2 promoter in vivo and in vitro indicates that the STP gene family indeed encodes transcription factors. The presence of a Leu3p binding site in the BAP2 promoter is required for full promoter activity on synthetic complete medium. The capacity of Leu3p to activate BAP2 transcription correlates with conditions that affect the level of alpha-isopropyl malate. The effect of a tup1 deletion on BAP2 transcription depends on SSY1. In an ssy1 strain, the phenotype of tup1 conforms to the well-established role of Tup1p as part of a repressor complex, but in the SSY1 strain deletion of TUP1 causes a decrease in transcription, indicating that Tup1p may also have an activating role at the BAP2 promoter. Our results thus suggest a complex interplay between several transcription factors in the expression of BAP2.

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Section: Discussionmentioning

confidence: 98%

“…Leu3p binds to its target DNA (5¢-CCGN 4 CGG-3¢) irrespective of the presence or absence of a-IPM, but at high a-IPM concentrations Leu3p is a strong transcriptional activator. At low a-IPM concentrations Leu3p can repress transcription (Wang et al 1997(Wang et al , 1999.…”

Section: Introductionmentioning

confidence: 99%

Transcriptional regulation of the Saccharomyces cerevisiae amino acid permease gene BAP2

et al. 2001

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“…Protease digestion assays indicated different accessibility of domains in Put3 (36). Two-hybrid assays between separated parts of Leu3 suggested interactions between central and C-terminal regions (14,15). Our attempts using similar two-hybrid approaches with War1 failed.…”

Section: Conserved Internal Domains Have Inhibitory Functions-mentioning

confidence: 95%

“…Put3 regulates proline utilization genes and is controlled by direct interaction with proline (12). Similarly, Leu3, a regulator of leucine biosynthesis, binds its target promoters and is only activated in the presence of the leucine precursor ␣-isopropyl malate (13)(14)(15).…”

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confidence: 99%

Weak Organic Acids Trigger Conformational Changes of the Yeast Transcription Factor War1 in Vivo to Elicit Stress Adaptation

Gregori

Schüller

Frohner

et al. 2008

Journal of Biological Chemistry

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The Saccharomyces cerevisiae zinc cluster regulator War1 mediates an essential transcriptional and adaptive response to weak organic acid stress. Here we investigate the mechanism of War1 activation upon weak acid stress. We identified several gain-of-function WAR1 alleles mapping to the central War1 region. These mutations constitutively increase levels of the plasma membrane ABC transporter Pdr12, the main War1 target mediating stress adaptation. Functional analysis of War1 reveals that the central region and its C-terminal activation domain are required for function. Notably, the native DNAbinding and dimerization domains appear dispensable for War1 activity, because they can be replaced by a LexA DNA-binding domain. Chromatin immunoprecipitation demonstrates elevated promoter affinity of activated War1, because its PDR12 promoter association increases upon stress. Hyperactive WAR1 alleles have constitutively high PDR12 promoter association. Furthermore, fluorescence resonance energy transfer of functional CFP-War1-YFP proteins also demonstrates conformational changes of stress-activated War1 in vivo. Our results suggest a mechanism whereby War1 activation is accompanied by conformational changes enhancing promoter association, thus initiating the adaptation process.Weak organic acids, such as potassium sorbate, calcium propionate, and sodium benzoate, are commonly used to preserve foods and beverages (1). In Saccharomyces cerevisiae, uncharged weak acids diffuse across the plasma membrane and dissociate inside the cell, causing intracellular acidification, free radical production, and oxidative stress (1), as well as inhibition of several metabolic processes (2). The yeast ABC transporter Pdr12 confers resistance to monocarboxylic acids with chain lengths up to C 7 (3, 4). Only organic acids but not other stresses cause a dramatic induction of PDR12 transcription and a concomitant increase of Pdr12 (4, 5). A physiological role of Pdr12 is suggested from its proposed ability to extrude amino acid catabolism by-products (6). Weak acid regulation of PDR12 occurs at the level of transcription and requires the Zn(II) 2 Cys 6 zinc cluster transcription factor War1, which recognizes and binds response elements within the PDR12 promoter (7). War1 is functionally conserved among several yeast species, including fungal pathogens. An orthologue mediating sorbate tolerance was identified in the distantly related human fungal pathogen Candida albicans (8), but the mechanism by which weak organic acids cause War1 activation is currently an open question.In S. cerevisiae, some 55 members of the binuclear zinc cluster regulator family exist (for a review see Ref. 9). Most exhibit a similar molecular architecture and domain organization. The N-terminal DNA-binding domain is followed by a coiled-coil dimerization domain, by an extended region of limited homology referred to as the "middle homology region" (MHR), 3 and finally by a short acidic stretch encompassing the C-terminal activation domain (10). Fungal zinc cluster pro...

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“…The concealment and exposure of XL Y are similar to a frequently observed feature of natural activation domains in which the hydrophobic residues that are vital for activator function are masked by either intramolecular or intermolecular interactions with other regulatory proteins (41)(42)(43)(44)(45)(46)(47)(48)(49)(50)(51)(52)(53). For example, in the structure of the p53 activation domain complexed with the inhibitor protein MDM-2, the hydrophobic residues that are known to play a critical role in activation are found buried in a hydrophobic cleft on the surface of MDM-2 (41,42).…”

Section: Discussionmentioning

confidence: 80%

Unraveling the Mechanism of a Potent Transcriptional Activator

Lü¹,

Rowe

Brennan

et al. 2005

Journal of Biological Chemistry

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Despite their enormous potential as novel research tools and therapeutic agents, artificial transcription factors (ATFs) that up-regulate transcription robustly in vivo remain elusive. In investigating an ATF that does function exceptionally well in vivo, we uncovered an unexpected relationship between transcription function and a binding interaction between the activation domain and an adjacent region of the DNA binding domain. Disruption of this interaction leads to complete loss of function in vivo, even though the activation domain is still able to bind to its target in the transcriptional machinery. We propose that this interaction parallels those between natural activation domains and their regulatory proteins, concealing the activation domain from solvent and the cellular milieu until it binds to its transcriptional machinery target. Inclusion of this property in the future design of ATFs should enhance their efficacy in vivo.

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Yeast Transcriptional Regulator Leu3p

Cited by 42 publications

References 33 publications

Transcriptional regulation of the Saccharomyces cerevisiae amino acid permease gene BAP2

Transcriptional regulation of the Saccharomyces cerevisiae amino acid permease gene BAP2

Weak Organic Acids Trigger Conformational Changes of the Yeast Transcription Factor War1 in Vivo to Elicit Stress Adaptation

Unraveling the Mechanism of a Potent Transcriptional Activator

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