Regulation of the yeast metallothionein gene

Gorman, Jessica A.; Clark, Philip E.; Lee, Ming Chou; Debouck, Christine; Rosenberg, Martin

doi:10.1016/0378-1119(86)90347-1

Cited by 41 publications

(29 citation statements)

References 16 publications

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“…Transcription of the yeast CUPI gene appears to be positively regulated by cis-acting upstream activation sequences (3,29), high concentrations of copper (3,18,29), and trans-acting transcriptional regulatory factors (9,12). A genetic approach was taken to begin to identify elements that regulate CUPI transcription in trails.…”

Section: Discussionmentioning

confidence: 99%

“…Biosynthesis of yeast MT is regulated at the level of transcriptional induction in response to high environmental copper levels (3,9,12,18,29). Although the yeast MT is able to bind to a variety of metals both in vivo and in vitro (5,32), copper is the only known inducer of CUPI gene transcription (3,18).…”

mentioning

confidence: 99%

“…transcriptional regulation are clear from the analysis of episomal CUP! promoter fusions in cup!A strains (9,12). (i) The CUPI protein itself is necessary to maintain low basal levels of transcription from the CUP!…”

mentioning

confidence: 99%

“…Because expression of heterologous MTs in cuplA strains restores low basal-level transcription to the CUP! promoter (9,30), CUP! likely regulates its expression by sequestering intracellular copper that would otherwise be available to putative copper-dependent transcription factors.…”

mentioning

confidence: 99%

“…likely regulates its expression by sequestering intracellular copper that would otherwise be available to putative copper-dependent transcription factors. (ii) Because episomal copies of the CUPI promoter are somewhat copper inducible in cupMA strains (9,30), a factor(s) other than CUP! protein must be involved in the copper-inducible transcription of CUPI.…”

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confidence: 99%

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ACE1 regulates expression of the Saccharomyces cerevisiae metallothionein gene.

Thiele¹

1988

Mol. Cell. Biol.

287

248

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Copper resistance in Saccharomyces cerevisiae is mediated, in large part, by the CUP1 locus, which encodes a low-molecular-weight, cysteine-rich metal-binding protein. Expression of the CUP1 gene is regulated at the level of transcriptional induction in response to high environmental copper levels. This report describes the isolation of a yeast mutant, ace1-1, which is defective in the activation of CUP1 expression upon exposure to exogenous copper. The ace1-1 mutation is recessive and lies in a genetic element that encodes a trans-acting CUP1 regulatory factor. The wild-type ACE1 gene was isolated by in vivo complementation and restores copper inducibility of CUP1 expression and copper resistance to the otherwise copper-sensitive ace1-1 mutant. Linkage analysis and gene deletion experiments verified that this gene represents the authentic ACE1 locus. ACE1 maps to the left arm of chromosome VII, 9 centimorgans centromere distal to lys5. The ACE1 gene appears to play a direct or indirect positive role in activation of CUP1 expression in response to elevated copper concentrations.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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ACE1 regulates expression of the Saccharomyces cerevisiae metallothionein gene.

Thiele¹

1988

Mol. Cell. Biol.

287

248

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Propagation of an amplifiable recombinant plasmid in Saccharomyces cerevisiae: flow cytometry studies and segregated modeling

Kd¹,

Je²,

Ratzkin³

et al. 1990

Biotech & Bioengineering

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Efficient expression of a foreign protein product by the yeast Saccharomyces cerevisiae requires a stable recombinant vector present at a high number of copies per cell. A conditional centromere yeast plasmid was constructed which can be amplified to high copy number by a process of unequal partitioning at cell division, followed by selection for increased copy number. However, in the absence of selection pressure for plasmid amplification, copy number rapidly drops from 25 plasmids/cell to 6 plasmids/cell in less than 10 generations of growth. Copy number subsequently decreases from 6 plasmids/cell to 2 plasmids/cell over a span of 50 generations. A combination of flow cytometric measurement of copy number distributions and segregated mathematical modeling were applied to test the predictions of a conceptual model of conditional centromere plasmid propagation. Measured distributions of plasmid content displayed a significant subpopulation of cells with a copy number of 4-6, even in a population whose mean copy number was 13.5. This type of copy number distribution was reproduced by a mathematical model which assumes that a maximum of 4-6 centromere plasmids per cell can be stably partitioned at cell division. The model also reproduces the observed biphasic kinetics of plasmid number instability. The agreement between simulation and experimental results provides support for the proposed model and demonstrates the utility of the flow cytometry/segregated modeling approach for the study of multicopy recombinant vector propagation.

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Evaluation of the Saccharomyces cerevisiae ADH2 promoter for protein synthesis

Lee

DaSilva

2005

Yeast

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The Saccharomyces cerevisiae ADH2 promoter (P ADH2 ) is repressed several hundredfold in the presence of glucose; transcription is initiated once the glucose in the medium is exhausted. The promoter can thus be utilized for effective regulation of recombinant gene expression in S. cerevisiae without the addition of an inducer. To evaluate this promoter in the absence of plasmid copy number and stability variations, the P ADH2 − lacZ cassette was integrated into the yeast chromosomes. The effects of medium composition, glucose concentration and cultivation time on promoter derepression and expression level were investigated. Maximum protein activity was obtained after 48 h of growth in complex YPD medium containing 1% glucose. The widely used S. cerevisiae GAL1 and CUP1 promoters both require the addition of an inducer [galactose and copper(II) ion, respectively] before regulated genes will be expressed. The strengths of these three different promoters were compared for cells containing one copy of an integrated lacZ gene under their control. The ADH2 promoter was superior for all induction strategies investigated.

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Regulation of the yeast metallothionein gene

Cited by 41 publications

References 16 publications

ACE1 regulates expression of the Saccharomyces cerevisiae metallothionein gene.

ACE1 regulates expression of the Saccharomyces cerevisiae metallothionein gene.

Propagation of an amplifiable recombinant plasmid in Saccharomyces cerevisiae: flow cytometry studies and segregated modeling

Evaluation of the Saccharomyces cerevisiae ADH2 promoter for protein synthesis

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